Rice cultivar designated 'cl261'.

(54) Title: RICE CULTIVATION CALLED CL261. (54) Title: RICE CULTIVAR DESIGNATED 'CL261'.

(57) Summary

The present invention relates to hybrid cultivars and derivatives of the rice cultivar named CL261. Hybrid rice seeds and plants produced by crossing cultivar CL261 with another rice cultivar. Other derivatives of rice cultivar CL261.

(57) Abstract

Hybrids and derived cultivars of the rice cultivar designated 'CL261' are disclosed. The invention relates to hybrid rice seeds and plants produced by Crossing the cultivar 'CL261' with another rice cultivar. The invention further relates to other derivatives of the rice cultivar 'CL261

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Mexican Property

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PATENT TITLE NO. 337901

Headlines):

Home:

Denomination:

Classification:

Inventor (s):

BOARD OF SUPERVISORS OF LOUISIANA STATE UNIVERSITY AND AGRICULTURE !. AND MECHANICAL COLLEGE LSU AgCenter, 104 Efferson Hall, Baton Rouge, Louislana, 70803, USA RICE CULTIVAR CALLED “CL261”.

lnt.CI.8: A01H1 / 02; A01H5 / 00

STEVEN D. LINSCOMBE

Number:

MX / a / 2012/003977

REQUEST

Intemaclomfc filing date October 2010

Country:

F ^us 1 ^us i

Validity: Twenty years

3 || JE

Expiration Date 7

PRIORITY

Date:

October 2009 October 4, 2010 &lt;·

Number:

61 / 249,641 PCT / US2010 / 05124Í

The reference patent is granted with fi

In accordance with article 23 of the rope, from the date of waste collection.

The undersigned of this title has been Industrial Property (Official Journal of

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of the Pi tion of fraction V. 6 ° Faction lll and 59 of the Industrial Property Law, this patent has a validity of twenty impi years and will be subject to the payment of the fee to maintain, according to articles 6 fractions lll and 7a bis 2 of laley of the reformed on 02.08.1994 10/25/1996, 12/26/1997, 1 05/1999, 26 ^ 11/2004, 06/16/2005, 01/25/2006, 0Λ5 / 2009,06 / 01 / 2010, 06/18/2010 / 06/26/2010, 01/27/2012 and 04/04/2012); articles 1, 3 | action V with foundation i Federation (DOF) 27Λ Indian a), 4 ° and 12 ° fracciom 01 ñrt &amp; sfó; izwrrísw:

Industrial Ladder (DOF 12/14/1999, laughed at the Organic Industrial Opiety (I of the Mexican Institute of Industrial Property (DOF 12/27/1999, amended on 10/10/2002, 07/29/2004, 08/04/2004 and 09/13/2007); 1, 3 and 5 subsection a) of the Agreement that delegates powers to the Deputy Directors General, Coordinator, Divisional Directors, Holders of the Regional Offices, Divisional Deputy Directors, Departmental Coordinators and other subordinates of the Mexican Institute of Industrial Property. (DOF 12/15/1999, amended on 02/04/2000, 07/29/2004, 08/04/2004 and 09/13/2007).

¹ and ^{111 of} Regulation of the Mexican Institute of Prop 72004 and

Re ^ arnent

Issue Date: March 28, 2016

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INDUSTRIAL

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The benefit of the filing date of October 4, 2010 of the international application PCT / US2010 / 051247 and the filing date of October 8, 2009 of the United States provisional patent application Series Number 61 / 249,641 are claimed under 35 USC Section 119 (e) in the United States and are claimed under applicable treaties and conventions in all countries.

TECHNICAL FIELD

This invention relates to a rice cultivar named "CL261", with hybrids thereof, and cultivars derived from the rice cultivar named "CL261".

TECHNICAL BACKGROUND

Rice is an ancient agricultural crop and remains one of the most important food crops in the world. There are two cultivated rice species: Oryza sativa L., Asian rice, and O. glaberrima Steud., African rice. Oryza sativa L. constitutes virtually all of the rice grown in the world, and is the species grown in the United States. There are three major rice-growing regions in the United States: the Mississippi Delta (Arkansas, Mississippi, Northeast Louisiana, Southeast Missouri), the Gulf Coast (Southwest Louisiana, Southeast Texas) and the Central Valley of California. See generally United States Patent No. 6911589.

Rice is a semi-aquatic crop that benefits under flooded soil conditions for part or all of the growing season. In the United States, rice is typically grown in its flooded area to optimize

<img file="MX337901B_D0005.tif" />

ι \; τ: γ. r; ,. ··. · The grain yield. Very clayey soil or loam with rigid impermeable layers approximately 30 cm below the surface are flexible soils for rice production, since they reduce the loss of water due to percolation. Rice production in the United States can be broadly classified as either dry planting or water planting. In the dry sowing system, rice is sown in a well prepared seed bed, with perforations for the grains or dispersing the seeds and incorporating them with a disc or a harrow. Moisture for seed germination comes from irrigation or rainfall. Another method for dry planting involves dispersing the seeds from an airplane in a flooded field, after which the field water is quickly drained. For ca sowing systems, when the plants have reached a sufficient size (a stage of four or five leaves), a permanent flood of shallow water, between 5 and 16 cm, is applied to the field during the rest of the growing season. Some of the rice is grown in unplanned production systems, without flooding.

A method of planting in water involves soaking the rice for 12-36 hours in order to start germination, and then dispersing the seeds with an airplane in a flooded field. Shoots emerge through a shallow flood, but it is also possible to drain the field water for a short period of time to improve the establishment of the shoots. Subsequently, a shallow flood is maintained until the rice approaches maturity. For dry sowing and water sowing production systems, the fields are drained when the crop is mature, and the rice is harvested 2-3 weeks later with large combines.

In rice development programs, those responsible for rice development typically use the same production systems that predominate

<img file="MX337901B_D0006.tif" />

in the region. Therefore, development with a drilling seed drill is typically used in those regions where rice is seeded drilling, and a development with water drilling is used in those regions where this system prevails.

Rice in the United States is classified into three primary marketing types based on grain size, grain shape, and endosperm composition: long-grain, medium-grain, and short-grain rice. Typical long grain rice cultivars in the US are dry and fluffy when cooked or boiled, while medium and short grain rice cultivars are moist and sticky when cooked. Medium grain rice is usually somewhat wider and somewhat shorter than long grain rice: it has a length / width ratio of between 2: 1 and 3: 1.

In the early part of the 1980s, approximately two-thirds of the Louisiana rice planting area was planted with medium-grain rice cultivars. Market demands and distribution of superior long-grain varieties (which, for example, have superior yields and superior disease resistance) resulted in a considerable decline in planting of medium-grain rice over the past 25 years, approximately. By the mid-1990s, Louisiana's rice planting area comprised 65 percent long-grain rice and 35 percent medium-grain rice. For the past 10 years or so, less than 5 percent of Louisiana's rice planting area has comprised medium-grain rice.

In recent years, long grain cultivars have become the leading cultivars, not only in Louisiana, but throughout the southern United States as well. However, the demand for the

<img file="MX337901B_D0007.tif" />

medium grain rice production in the region, and recently this demand has increased substantially. Although various medium grain rice cultivars lacking herbicide resistance are available in the region, such as "Jupiter", "Neptune" and "Bengal", no herbicide resistant rice cultivars have been distributed.

There is an unmet need for herbicide resistant medium grain rice cultivars, particularly cultivars that are well adapted to the prevailing growing conditions in the southern United States. Medium grain rice is more common in California, but in the southern United States, medium grain rice differs somewhat from that grown in California. Additionally, weed known as “red rice” has become a substantial problem in medium-grain rice fields in the southern United States. Medium grain rice cultivars that are resistant to herbicides have not been distributed. There is a particular need for herbicide resistant medium grain rice cultivars.

Although specific development objectives vary to some extent across regions, increasing performance is a primary objective in all programs. Grain yield depends in part on the number of panicles per unit area, the number of fertile panicles per panicle, and the weight of the grains per floret. Increases in any or all of these components can help improve yields. Each of these components has a certain hereditary variation, and those in charge of development can make a direct or indirect selection in favor of any of them.

There are numerous steps in the development of any desirable novel plant germplasm. The development of a plant begins with I analysis and

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definition of the problems and weaknesses of the current germplasm, the establishment of program objectives and the definition of specific development objectives. The next step involves selecting (or generating) a germplasm that has the characters that meet the program's objectives. Frequently, the goal is to combine an improved character set from two or more ancestral germplasm lines into a single variety. These traits may include improved seed yield, resistance to diseases or insects, improved stems and roots, tolerance to low temperatures, and improved agronomic characteristics or grain quality.

The choice of development and selection methods depends on the mode of reproduction of the plant, the hereditary character of the one or more characters that it is desired to improve and the type of seeds that are used in the commercial field (for example, hybrids Fi compared to cultivars of pure or inbred lines). For highly hereditary traits, it may occasionally be effective to select the top individual plants that are evaluated at a single location. However, for traits that are inherited by complex means or to a limited extent, selection is often based on the mean values ​​obtained from repeated evaluations of related plant families. Selection methods include pedigree selection, modified pedigree selection, mass selection, recurring selection, and combinations of these methods.

The complexity of inheritance influences the choice of development method. Backcross development is used to transfer a gene or a few genes favorable for a highly hereditary character to a desirable cultivar. This approach has been widely used to develop disease resistant cultivars. Various recurrent selection techniques are used to enhance traits that are quantitatively inherited across numerous genes. The use of recurrent selection in self-pollinated crops depends on the ease of pollination, the frequency of successful hybrids at each pollination, and the amount of hybrid offspring from each successful cross.

Promising lines during advanced stages of development are thoroughly evaluated and compared to appropriate benchmarks in settings representative of desired business areas, typically for three years or more. The best lines become candidates for new commercial cultivars; Those that still have deficiencies in a few characters can be used as parents to produce new populations to continue with the selection.

These processes, which ultimately result in the commercialization and distribution of cultivars or hybrids, typically take 8 to 12 years from the time of the first cross. They may also be based on the development of improved crossing lines as precursors (which may introduce additional delay). The development of novel cultivars and hybrids is a laborious process for which precise planning and efficient use of resources are required. There is no assurance of a successful outcome.

A particularly difficult task is the identification of those individual plants that are indeed superior in genetic terms. The phenotype of a plant is the result of a complex interaction between genetics and the environment. One method of identifying a genetically superior plant involves observing its performance compared to that of other plants.

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experimental and with that of a widely cultivated reference cultivar, which has been developed in an Identical environment. Repeated observations can be made at multiple locations to help better estimate their genetic value.

The goal of rice development is to develop unique, superior rice cultivars and hybrids. Initially, growers select and cross two or more progenitor lines, followed by self-pollination and selection, in order to produce many novel genetic combinations. Cultivators can generate billions of different genetic combinations through crossing, self-crossing, and mutational development. Traditional cultivators do not have direct control at the molecular level. So two traditional growers working independently will never develop the same line, or even very similar lines, with the same characters.

Every year, plant managers select germplasm to take it to the next generation. Germplasm is cultivated under different geographical, climatic and soil conditions. Additional selections are then made during the growing season and towards the end of the growing season. The resulting cultivars (or hybrids) and their characteristics are inherently unpredictable. This is because the selection made by traditional growers takes place in unique environments, without control at the molecular level, and with the potential generation of billions of different possible genetic combinations. Crop managers cannot predict the final resulting line, except possibly in a very broad and generic way. Furthermore, the same cultivator may not produce the same cultivar twice, even if it starts with the same cultivars.

<img file="MX337901B_D0010.tif" />

parent lines and will use the same selection techniques. This uncontrollable variation results in substantial effort and cost in developing new superior rice cultivars (or hybrids) and prevents prediction of the outcome of each novel cultivar (or hybrid).

The selection of the superior hybrid crosses is made in a slightly different way. Hybrid seeds are typically produced by manual crossing between selected parents with male fertility, or by using systems with genetic male sterility. These hybrids typically undergo selection in favor of individual gene-based characters that unambiguously indicate that a plant is indeed a Fi hybrid that has inherited the characters from both presumed parents, particularly the male parent (since rice normally self-fertilizes). These characters could include, for example, a plant of a semi-dwarf type, pubescence, the color of the ridges or of the apiculus. Other data on the progenitor lines, as well as the phenotype of the hybrid, influence the decisions of the cultivators related to the continuation with a particular hybrid cross or with an analogous cross, by means of unrelated progenitor lines.

Pedigree development and recurrent selection development methods are occasionally used to develop cultivars from developing populations. In these development methods, desirable traits from two or more cultivars or other germplasm sources are combined in developmental reserves, from which cultivars can be developed by self-pollination and by selection of the desired phenotypes. New cultivars are evaluated to determine their commercial potential.

<img file="MX337901B_D0011.tif" />

Pedigree development is often used to improve self-pollinated crops. Two parents possessing favorable complementary characters interbreed to produce Fi plants. An F2 population is produced by autocrossing one or more F1 hybrids. The selection of the superior individual plants can start in the F2 generation (or in a later generation). Then, from the F3 generation (or a subsequent generation), the individual plants are selected. Repeated evaluation of the panicle rows of the selected plants can begin in the F4 generation (or in another subsequent generation), both to fix the desired characters and to improve the effectiveness of the selection of those characters that are inherited to a reduced extent. . When an advanced level of inbreeding has been reached (for example, F₆ or F₇), the best lines or mixtures of lines with similar phenotypes are evaluated to determine if it is possible to distribute them as new cultivars.

Recurrent mass selection methods can also be used to improve populations of crops that are self-pollinated or cross-pollinated. A genetically variable population of heterozygous individuals is identified or created by crossing several different parents. The best progeny plants are selected on the basis of their individual superiority, outstanding progeny, or excellent combining ability. The selected plants are crossed to produce a new population, with which development continues in additional cycles.

Backcross development is often used to transfer genes of a single and frequent inheritance trait to a desirable cultivar or homozygous inbred line, which is the recurrent parent. The source of the character to be transferred is called the donor parent. Ideally, the

<img file="MX337901B_D0012.tif" />

plant should have the attributes of the recurring parent (eg cultivar) and the new desired character transferred from the donor parent. After the initial crossover, those individuals possessing the desired donor phenotype (eg, disease resistance, insect resistance, herbicide tolerance) are selected and subjected to repeated crosses with the recurring parent (backcrossed).

Strictly speaking, the single-seed offspring procedure refers to planting a segregating population, harvesting a one-seed sample per plant, and using the one-seed sample to sow the next generation. When the population has advanced from generation F₂ to the desired level of inbreeding, the representative plants of the derived lines can be traced to the different individuals F₂. The number of plants in a population is reduced in each generation due to the inability of some seeds to germinate or the inability of some plants to produce at least one seed. As a result, not all F plants₂in the population from which a sample was originally taken, they will be represented in the progeny over the generations.

In a multi-seed procedure, the grower harvests one or more seeds from each plant in the population and threshes them to form a batch. Part of the lot is used to seed the next generation, and part is placed in reserve. The procedure is known as a modified single seed offspring technique or as a pod lot technique. The multiple seed procedure has been used to save labor at harvest. It is considerably faster to thresh the panicles by machine than to remove one seed from each plant by hand, as is the case with the single seed procedure. The multi-seed procedure is also possible

<img file="MX337901B_D0013.tif" />

sow all the seeds of a population in each generation of inbreeding. Enough seeds are harvested to compensate for plant loss due to lack of germination or inability to produce seeds.

Other common and less common development methods are known and used in the art. See, for example, RW Allard, Principies of Plant Breeding (John Wiley and Sons, Inc., New York, New York, 1967), NW Simmonds, Principies of Crop Improvement (Longman, London, 1979), J. Sneep et al ., Plant Breeding Perspectives (Pudoc, Wageningen, 1979), and WR Fehr, Principles of Cultivar Development: Theory and Technique (Macmillan Pub., New York, New York, 1987).

By means of an appropriate evaluation, it would be possible to detect any significant error and establish the level of superiority or improvement compared to current cultivars. In addition to superior performance, there is a demand for a new cultivar or hybrid that is compatible with industry guidelines or that can create a new market. The introduction of a new cultivar or hybrid may represent additional costs for the seed producer, for the cultivator, for the processor and for the consumer, for example, in specialized advertising and marketing, in seed production practices and commercial changes and in the use of new products. During pre-distribution evaluation of a new cultivar or hybrid, costs in research and development should be taken into account, as well as the technical superiority of the cultivar or final hybrid.

In recent years, a few herbicide-tolerant rice varieties and hybrids have been successfully introduced to the market. See US Patent Nos. 5545822, 5736629, 5773703, 5773704, 5952553, 6274796, 6943280, 7019196, 7345221, 7399905 and 7495153, International Applications.

<img file="MX337901B_D0014.tif" />

of published patents WO 00/27182 and WO 01/85970 and published US patent application 2007/0061915. These herbicide-tolerant rice plants exhibit herbicide resistance or tolerance that would normally inhibit the growth of the rice plants. Therefore, growers can now control weeds that were previously difficult to control in rice fields, including “red rice”. “Red rice” is a weed related to cultivated rice that had previously been difficult to control because it actually belongs to the same species as cultivated rice. Only recently, with the distribution of herbicide-tolerant rice varieties, has it been possible to control red rice with herbicides in fields where commercial rice was being grown simultaneously. Currently, there are only a very limited number of commercially available herbicide tolerant cultivars and hybrids. There is a continuing need for new herbicide tolerant cultivars and hybrids, i.e., rice plants where not only is a desired herbicide tolerance phenotype expressed, but they also possess other agronomically desirable characteristics. These additional cultivars and hybrids will provide rice growers with greater flexibility in planting and crop management.

DESCRIPTION OF THE INVENTION

A novel herbicide-resistant rice cultivar of high yield, early maturation, short stature and medium grains was discovered which was named “CL261”. This invention also relates to methods of producing a novel or hybrid variety comprising making one or more "CL261" rice variety crosses with another rice line. By

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Accordingly, all methods where the "CL261" variety of rice is used will be aspects of this invention, including backcrossing, hybrid production, population crossing, and other development methods where "CL261" is employed. Hybrid plants that are produced using the rice variety "CL261" as parent will also be within the scope of this invention. Optionally, it will be possible to produce any parent with male sterility, by routine manipulation of cytoplasmic or other factors, in accordance with procedures known in the art.

In another embodiment, converted plants with a single "CL261" gene are provided in this invention. The transferred individual gene can be a dominant or recessive allele. Preferably, the transferred individual gene confers a character such as resistance to insects or to one or more diseases of bacterial, fungal, or viral origin, male fertility or sterility, improved nutritional quality, improved processing qualities, or an additional source of resistance to herbicides. The individual gene may be a natural rice gene or a transgene introduced through genetic engineering procedures known in the art. The individual gene can also be introduced by means of traditional backcross or genetic transformation procedures known in the art.

In another embodiment, regenerable cells for use in tissue culture of a "CL261" rice plant are provided in this invention. Tissue culture can allow to regenerate plants with physiological and morphological characteristics of a "CL261" rice plant, and also to regenerate plants with a genotype substantially identical to that of the "CL261" rice plant. Procedures for cultivating rice tissue are known in the art. The

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Regenerable cells in tissue culture can derive from sources such as embryos, protoplasts, meristematic cells, corns, poln, leaves, anthers, root tips, flowers, seeds, panicles or the stems. Furthermore, regenerated rice plants are provided in the invention from such tissue culture.

In other embodiments, a method of controlling weeds in the vicinity of rice is provided in the present invention. The method involves contacting the rice with a herbicide, where said rice belongs (a) to the variety “CL261” or (b) to a hybrid, a derivative or the “CL261” program where the resistance characteristics are expressed to the imidazolinone herbicide of "CL261".

In some embodiments, the herbicide is an imidazolinone herbicide, a sulfonylurea herbicide, or a combination of these.

In one embodiment, the rice is a rice plant and said contacting comprises applying the herbicide in the vicinity of the rice plant.

In another embodiment, the herbicide is applied to the weeds in the vicinity of the rice plant.

In still other embodiments, the rice is a rice seed and such contacting comprises applying the herbicide on the rice seed.

In some embodiments, a method of treating rice is provided in the present invention. The method comprises contacting the rice with an agronomically acceptable composition, where said rice belongs (a) to the variety "CL261" or (b) to a hybrid, a derivative or progeny of "CL261" where the characteristics of resistance to the herbicide imidazolinone of "CL261".

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INSTITUTO .MEXICANO

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INDUSTRIAL

In one embodiment, the agronomically acceptable composition comprises at least one agronomically acceptable active ingredient.

In another embodiment, the agronomically acceptable active ingredient is selected from the group consisting of fungicides, insecticides, antibiotics, compounds that enhance stress tolerance, growth promoters, herbicides, molluscicides, rodenticides, animal repellents and combinations of these.

DEFINITIONS

The following definitions apply to the entire specification and claims, unless the context clearly arises otherwise.

"Days until reaching 50% of ear". Average number of days from planting to the day that 50% of the panicles are at least partially extended through the leaf sheath. It is a measure of maturity.

"Yield of the grains". Grain yield is measured in pounds per acre, with a humidity of 12.0%. Grain yield depends on a number of factors, including the number of panicles per unit area, the number of fertile florets per panicle, and the weight of the grains per floret.

"Tipping percentage". Tipping is a subjectively measured rating: it is the percentage of plant stems that completely bow or fall to the ground before harvest.

"Length of the grains (L)". The length of a grain of rice, or the average length, measured in millimeters.

"Grain width (W)". The width of a grain of rice, or the average width, measured in millimeters.

<img file="MX337901B_D0019.tif" />

"Length / width ratio (L / W)". This ratio is determined by dividing the average length (L) by the average width (W).

"Weight of 1000 grains". The weight of 1000 grains of rice, measured in grams.

"Humidity at harvest". The percentage of moisture in the grains at the time of harvest.

"Height of the plant". The height of the plant in centimeters, measured from the soil surface to the end of the extended panicle at harvest.

"Percentage of apparent amylose". The percentage of starch in the form of amylose in the endosperm of ground rice. The apparent amylose percentage is an important characteristic of the grains that affects the cooking behavior. Conventional long grains contain between 20 and 23 percent amylose. Long Rexmont-type grains contain 24 to 25 percent amylose. Short and medium grains contain between 13 and 19 percent amylose. Waxy rice contains zero percent amylose. Like most characteristics of rice, amylose values ​​depend on ambi nt. "Apparent" refers to the procedure used to determine amylose, which may also comprise measuring some of the long-chain amylopectin molecules that bind to some of the amylose molecules. In fact, these amylopectin molecules act similarly to amylose in determining the characteristics of hard or soft cooking.

"Alkali dispersion value". An index that reflects the extent of disintegration of crushed rice grains when contacted with a dilute alkaline solution. It is an indicator of the temperature of

<img file="MX337901B_D0020.tif" />

gelatinization. Conventional long grains have an alkali dispersion value of between 3 and 5 (an intermediate gelatinization temperature).

"Maximum viscosity". The maximum viscosity that is reached during heating when a standard, instrument-specific protocol is applied to a defined suspension of rice flour in water.

"Minimum viscosity". The lowest viscosity after the maximum viscosity, which normally occurs when the sample begins to cool.

"Final viscosity". The viscosity at the end of the test or of the cold paste.

"Degradation". The maximum viscosity minus the viscosity of the hot paste.

"Regression". Regression 1 is the final viscosity minus the minimum viscosity. Regression 2 is the final viscosity minus the maximum viscosity.

"Viscosity RVA". Viscosity measured with a fast viscosity analyzer, which is a novel but widely used laboratory instrument to examine the viscosity of a paste or the thickening capacity of minced rice during the cooking process.

"Viscosity of hot pasta". The measured viscosity of a suspension of rice flour / water after heating to 95 ° C. The lower values ​​are indicative of types of rice that are softer and stickier in cooking.

"Viscosity of cold pasta". The measured viscosity of a rice flour / water suspension after heating to 95 ° C and cooling it uniformly to 50 ° C. Values ​​less than 200 are indicative of types of rice that are softer in cooking.

Allele. An allele is any one or more alternative forms of the same gene. In a diploid cell or organism, the two alleles of a given gene occupy corresponding loci in a pair of homologous chromosomes.

<img file="MX337901B_D0021.tif" />

"Backcross". Backcrossing is a process in which the developer repeatedly crosses hybrid progeny with one or more of the parents, for example, crossing a first generation Fi hybrid with one of the parent genotypes of the Fi hybrid, and then crossing a second generation F2 hybrid with the same parent genotype, and so on.

"Essentially all of the physiological and morphological characteristics." A plant that has "essentially all of the physiological and morphological characteristics" of a specific plant refers to a plant that has the same general physiological and morphological characteristics, except for characteristics derived from a particular converted genome.

"Loci of a quantitative character (QTL). Quantitative Character Loci (QTL) refer to genetic loci that control characters that can be measured numerically to some degree; generally these are characters that have a continuous distribution.

"Regeneration". Regeneration refers to the development of a plant from a tissue culture.

"Converted with a single gene (conversion)". Plants converted with a single gene (conversion) include plants that are grown by a backcross where essentially all of the desired morphological and physiological characteristics of the parent variety are recovered, while retaining the individual gene transferred to the variety through crossing and backcrossing. The term may also refer to the introduction of a single gene by means of genetic engineering procedures known in the art.

<img file="MX337901B_D0022.tif" />

WAYS TO PRACTICE THE INVENTION “CL261” is a very high yielding rice variety, early maturity, small stature and medium grains that, by virtue of their common ancestors, contains the same herbicide resistance genes found in the "CL161". The pedigree of this line is "Bengal" / "CL161". "CL161" is an imazetapir resistant mutant derived from the Cypress variety. See US Patent No. 7019196. "CL261" is highly resistant to imidazolinone herbicides, including, without limitation, imazetapir and imazamox. “CL261” has light grains, without much chalk. Its prominent grains are similar in size to those of the medium grain varieties "Neptune" and "Bengal".

"CL261" was originally selected from a "Bengal7" CL161 "cross, which was conducted at the Louisiana State University Rice Research Station in Crowley, Louisiana. The line originated as batch F₅ from a single set of progeny. "CL261" is small in stature and modestly resistant to tipping. It presented an average height of 37 inches during the two years of evaluation, compared to 37 inches and 33 inches for cultivars "CL151" and "CL131", respectively. “CL261” takes an average of 81 days from emergence to reach 50% gleaning, compared to 79 days for “CL151” and “CL131”. The leaves, motto, and blade of CL261 are glabrous. The spikelets are straw colored. The apicule has a light purple color at the time of bolting, which disappears as the grains mature. The beans are not aromatic. "CL261" is susceptible to sheath blight and blight, and is moderately susceptible to physiological vaneum.

After the initial crossing, the line was harvested and selected during the first generations to achieve phenotypic superiority in

<img file="MX337901B_D0023.tif" />

characteristics such as the architecture of a ground floor, the shape and uniformity of the grains, the vigor of the seeds, the number of reeds and the size of the grains. In later generations (as the seeds increased), the line was selected in favor of uniformity and purity, both within and between the rows of panicles. In principle, the variants taken from fields where the “CL261” seeds had been increased were taller or later maturing plants. Other variants included any combination of the following characteristics: earlier and shorter leaf pubescence, long grains, golden peel, lighter colored leaves, and variegated leaves. The overall incidence of variants was 1 per 5,000 plants. Eventually, the founder rice seeds were grown from the F7 generation. Seeds from the F5, F6 and F7 generations were introduced into a program to evaluate experimental lines, and were also evaluated at various locations in the rice growing areas of Louisiana. "CL261" was observed to be stable for at least three generations.

During evaluation in various trials at multiple locations in Louisiana over a three-year period, the average overall grain yield was 7,871 lbs / acre for “CL261”, compared to 7,700 lbs / acre for “CL131” and 8,683 lbs / acre for “CL151”.

DESCRIPTIVE INFORMATION OF THE VARIETY

The rice cultivar “CL261” has been observed to possess the following morphological and other characteristics, based on averages of evaluations conducted at multiple locations in the state of Louisiana over three growing seasons. For comparative purposes, the data of the varieties “CL131” and “CL151” are provided.

<td>Character</td><td colspan="3">Performance</td><td colspan="2">Ijr. _____ · ς γ &lt;· \ ΐ ...-.:,../.3 Quantity -tfe-- evaluated nes</td>

<td></td><td>"CL261"</td><td>"CL131"</td><td colspan="2">"CL151"</td><td></td>

<td>Average crop yield (pounds / acre)</td><td> 7871</td><td> 7700</td><td colspan="2"> 8683</td><td> 6</td>

<td>Yield of the shoots of the crop (pounds / acre)</td><td></td><td></td><td colspan="2"></td><td></td>

<td>Crushing performance of whole rice (%)</td><td> 67,4</td><td> 65,6</td><td colspan="2"> 63,8</td><td> 5</td>

<td>Total crushing performance of rice (%)</td><td> 70,8</td><td> 71,3</td><td colspan="2"> 70,0</td><td> 5</td>

<td>Vigor of suckers (rating subjective vigor of shoots on a scale of 1 to 9, where smaller values represent higher levels of vigor)</td><td> 4</td><td> 4</td><td colspan="2"> 4</td><td> 5</td>

<td>Average plant height (inch)</td><td> 37</td><td> 33</td><td colspan="2"> 37</td><td> 6</td>

<td>Average number of days until reach 50% spike</td><td> 81</td><td> 79</td><td colspan="2"> 79</td><td> 6</td>

<td>Reaction to the brown stain narrow (Cercospora janseana) (0 = very resistant, 9 = very</td><td></td><td> 4,8</td><td colspan="2"> 3,8</td><td></td>

<img file="MX337901B_D0024.tif" />

<td>susceptible)</td><td></td><td></td><td></td><td></td>

<td>Carbon reaction of the leaf (Entyloma oryzae) (0 = very resistant, 9 = very susceptible)</td><td></td><td></td><td></td><td></td>

<td>Reaction to rice worms (Helminthosporium oryzae) (0 = very resistant, 9 = very susceptible)</td><td></td><td></td><td></td><td></td>

<td>Reaction to rust panicles (B. gladioli) (0 = very resistant, 9 = very susceptible)</td><td></td><td> 6,3</td><td> 5,3</td><td></td>

<td>Reaction to black scabies (Rhizoctonia sotaní) (0 = very resistant, 9 = very susceptible)</td><td></td><td> 7,5</td><td> 6,5</td><td></td>

<td>Reaction to physiological vane (0 = very resistant, 9 = very susceptible)</td><td> 4,9</td><td> 6,7</td><td> 7,0</td><td> 2</td>

<td>Character</td><td colspan="3">Performance</td><td>Cant ad d evaluates gifts</td>

<td></td><td>"CL261"</td><td>"Jupiter"</td><td>Neptun and"</td><td></td>

<td>Rough length (mm)</td><td> 7,40</td><td> 7,87</td><td> 7,95</td><td></td>

<td>Gross width (mm)</td><td> 3,22</td><td> 3,20</td><td> 3,25</td><td></td>

ΜΡΪ

<td>Raw L / W Ratio</td><td> 2,30</td><td> 2,46</td><td> 2,45</td><td></td>

<td>Brown Serving Length (mm)</td><td> 5,75</td><td> 5,85</td><td> 5,97</td><td></td>

<td>Width of brown portion (mm)</td><td> 2,89</td><td> 2,86</td><td> 2,85</td><td></td>

<td>Portion L / W ratio Brown</td><td> 1,99</td><td> 2,05</td><td> 2,09</td><td></td>

<td>Length after completion of crushing (mm)</td><td> 5,31</td><td> 5,47</td><td> 5,62</td><td></td>

<td>Width once finished crushing (mm)</td><td> 2,66</td><td> 2,68</td><td> 2,55</td><td></td>

<td>L / W ratio once finished crushing</td><td> 2,00</td><td> 2,04</td><td> 2,20</td><td></td>

Cane

Angle (degrees from the perpendicular after flowering): erect (less than 30 °)

Length: 94cm (same as CL151, 10cm more than CL131)

Internode color (after flowering): green Strength (tip resistance): moderately large

Flag leaf (at maturity)

Length: 21cm

Width: 10mm

Pubescence: glabrous

Blade angle (after bolting): intermediate

Border color (on the spike): dark green

Color of basal leaf sheath (in the spike): v rde

<img file="MX337901B_D0025.tif" />

Ligule

Length: 2cm

Color (late vegetative state): white

Shape: cracked

Collar color (late vegetative state): pale green

Atrium color (late vegetative state): pale green Panicle

Length: 19 cm

Type: intermediate

Secondary branching: abundant

Exserción (near maturity):&gt; 90%

Flattening: (&lt;5%)

Plow Susceptibility: Intermediate Grains (Spikelets)

Edges (once the spike is complete): short and partial

Apiculum color (at maturity): purple

Apiculum color (once the spike is complete): purple

Stigma Color: White

Color of lemma and palea (at maturity): straw

Lemma pubescence and palea: glabrous

Spikelet sterility (at maturity): (&lt;10% = highly fertile) Grains (seeds)

Color of seed coat (germ): light brown

Endosperm type: non-glutinous (non-waxy)

Endosperm translucency: transparent

Presence of chalk in the ndosp rma: (scarce, less than 10% of the sample)

Α HA. jx.

L ·., I, ·· .. i&gt; i: ·. · - _ ;. ·· '&lt;- ^ 3

Aroma: absent

Shape class (length / width ratio)

Paddy - medium (2.3-3.3) (i.e. 2.30)

Brown - short (2.0 or less) (i.e. 1.99)

Crushed - medium (2.0-2.9) (i.e. 2.00)

<td>Shape of grain</td><td>Length</td><td>Width</td><td>Thickness</td><td>L / W</td><td>1000 grain</td>

<td></td><td>(mm)</td><td>(mm)</td><td>(mm)</td><td>Relationship</td><td>(grams)</td>

<td></td><td></td><td></td><td></td><td></td><td></td>

<td>Rice paddy</td><td> 7,40</td><td> 3,22</td><td> 1,99</td><td> 2,30</td><td> 24,6</td>

<td>Brown</td><td> 5,75</td><td> 2,89</td><td> 1,77</td><td> 1,99</td><td> 20,1</td>

<td>Crushed</td><td> 5,31</td><td> 2,66</td><td> 1,70</td><td> 2,00</td><td> 16,9</td>

Shredding quality (% of husks): 20 10 Shredding yield (% of white rice grains (in the ear) in the raw rice): 66-71

Proteins (NIR): 7.5%

Amylose: 18.9%

Alkali dispersion value: 7.0 (with 1.7% KOH solution)

Gelatinization temperature type: intermediate

Low temperature resistance

Vigor in germination and planting: medium Flowering (fertility of the spikelets): high ΐ, w,

Sprig vigor not related to low temperatures

Vigor: high

Insect resistance. Susceptible to the aquatic rice weevil (Lissorhoptrus oryzophilus) and the rice odor bug (Oebalus pugnax).

The variety is resistant to imidazolinone herbicides. The herbicide resistance profile is essentially the same as that of “CL161”, by virtue of common ancestors. Herbicide tolerance makes it possible to use “CL261”, its hybrids and its varieties derived with rice technology and Clearfield ™ herbicides, which include, among others, imazetapir and imazamox, for selective weed control, including red rice. See generally US Patent No. 6943280.

Herbicide tolerance and susceptibility characteristics The variety has tolerance to some herbicides and is susceptible to some herbicides that normally inhibit the growth of rice plants. Amongst others, the herbicide tolerance and susceptibility characteristics of “CL261” include those listed below, or at least are expected to include them. In some cases, these characteristics are based on actual observations to date, and in other cases, they reflect ancestor-based conjectures in common with “CL161”:

In "CL261" is expressed a mutant acetohydroxy acid synthetase whose enzymatic activity is directly resistant to levels of a herbicide-effective imidazolinone that would normally cause inhibition;

"CL261" is resistant to each of the following imidazolinone herbicides, niv I s in which imidazolinone herbicides would normally inhibit the

<img file="MX337901B_D0026.tif" />

growth of a rice plant: imazetapir, imazapic, imazaquin, imazamox and imazapir;

"CL261" is resistant to each of the following sulfonylurea herbicides, at levels at which sulfonylurea herbicides would normally inhibit the growth of a rice plant: nicosulfuron, metsulfuron methyl, tifensulfuron methyl and tribenuron methyl;

"CL26r is sensitive to each of the following sulfonylurea herbicides, at levels at which sulfonylurea herbicides would normally inhibit the growth of a rice plant: methyl sulfometuron, ethyl chlorimuron, and rimsulfuron.

This invention also relates to methods for producing a rice plant, comprising crossing a first parent rice plant with a second parent rice plant, where the first rice plant or the second rice plant are line rice plants "CL261". Furthermore, the first parent rice plant and the second parent rice plant may be from the cultivar "CL261", although it is preferable that one parent is different. Methods for using the cultivar "CL261" are part of this invention, and include crossbreeding, autocrossing, backcrossing, hybrid development, crossbreeding with populations, the other development methods previously described in this specification and other development methods to be known to those skilled in the art. Any plant that is produced using cultivar "CL261" as a parent or ancestor will be within the scope of this invention. The other parents or the other lines used in these development programs can be any one of a large number of varieties, cultivars, populations or lines

IMPI

MEXICAN INSTITUTE OF PROPERTY

INDUSTRIAL

<img file="MX337901B_D0027.tif" />

experimental rice, including other sources of rice germplasm known in the art.

For example, this invention encompasses methods for producing a first generation hybrid rice plant, which comprises crossing a first parent rice plant with a second parent rice plant, where the first parent rice plant or the second parent rice plant they are “CL261”. Furthermore, this invention also relates to methods for producing a hybrid rice line derived from "CL261", comprising crossing "CL261 with a second rice plant and cultivating progeny seeds. The crossing and cultivation steps can be repeated any number of times. Development methods where the “CL261” rice line is used are considered part of this invention, and include not only backcrossing and hybrid production, but also self-crossing, population crossbreeding, and other development methods known in the technique.

Optionally, either parent in the cross, either "CL261" or the other parent, may be produced with male sterility, using procedures known in the art.

Other embodiments of the invention

As used herein, the term "plant" includes plant cells, plant protoplasts, plant cells in tissue cultures from which rice plants, plant calluses, plant groups and intact plant cells can be regenerated in plants or plant parts such as pollen, flowers, embryos, ovules, seeds, pods, leaves, stems, anthers, and the like. Accordingly, another aspect of this invention comprises providing cells that, by developing and differentiating, produce a cultivar that

<img file="MX337901B_D0028.tif" />

it has essentially all of the physiological and morphological characteristics of "CL261".

Procedures for transforming desired structural genes, expressing them, and culturing cells are known in the art. Also, as is known in the art, rice can be transformed and regenerated to obtain complete plants containing and expressing the desired genes under appropriate regulatory control. General descriptions of expression vectors, reporter genes, and transformation protocols for plants can be found, for example, in Gruber et al., "Vectors for Plant Transformation, in Methods in Plant Molecular Biology &amp; Biotechnology", Glich et al. (editors, pp. 89-119, CRC Press, 1993). For example, expression vectors and gene cassettes with the GUS reporter gene are available from Clone Tech Laboratories, Inc. (Palo Alto, Calif.), And expression vectors and gene cassettes with the luciferase reporter gene are available from Promega Corp. (Madison, Wis.). General methods for growing plant tissues are provided, for example, in Maki et al., "Procedures for Introducing Foreign DNA into Plants", in Methods in Plant Molecular Biology &amp; Biotechnology, Glich et al., (Editors, pp. 67-88 CRC Press, 1993), in Phillips et al., “Cell-Tissue Culture and In-Vitro Manipulation”, in Corn &amp; Corn Improvem nt, 3^to edition, and in Sprague et al., (editors, pp. 345-387), American Society of Agronomy, Inc., 1988. Methods for introducing expression vectors into plant tissue include direct infection or concurrent culture of plant cells with Agrobacterium tumefaciens, Horsch et al., Science, 227: 1229 (1985). Descriptions of Agrobacterium vector systems and methods for effecting Agrobacterium-mediated gene transfer are provided in Gruber et al., Supra.

. 1

<img file="MX337901B_D0029.tif" />

Useful methods include, without limitation, expression vectors that are introduced into plant tissues using a direct gene transfer method, such as microprojectile-mediated administration, DNA injection, electroporation, and the like. More preferably, the expression vectors are introduced into plant tissues using microprojectile-mediated administration, in combination with transformation with a blolistic device or Agrobacterium-mediated. Transformed plants obtained with the germplasm of "CL261" are to be within the scope of this invention.

Regenerated rice plants from a tissue culture of the "CL261" variety, as well as hybrid plants, are also provided in the present invention. As is known in the art, tissue culture can be used for in vitro regeneration of a rice plant. For example, see Chu, QR et al. (1999) "Use of brldglng parents with hlgh anther culturablllty to ¡mprove plant regeneratlon and breedlng valué ¡n rice", Rice Biotechnology Quarterly, 38: 25-26, Chu, QR et ai, "A novel plant regeneration medium for rice anther culture of Southern US crosses", Rice Biotechnology Quarterly, 35: 15-16 (1998), Chu, QR et al., "A novel basal medium for embryogenlc callus inductlon of Southern US crosses ”, Rice Biotechnology Quarterly, 32: 19-20 (1997), and Oono, K.,“ Broadening the Genetic Varlabillty By Tlssue Culture Methods ”, Jap. J. Breed., 33 (Suppl. 2), 306-307 (1983). Therefore, another aspect of this invention comprises providing cells that, when developing and differentiating, produce rice plants that have all or essentially all of the physiological and morphological characteristics of the variety "CL261".

Unless the context clearly arises otherwise, references in the specification and claims to "CL261" must also include

<img file="MX337901B_D0030.tif" />

the genetic conversions of “CL261” to obtain male sterility, ^ other sources of resistance to herbicides, resistance to diseases of bacterial, fungal or viral origin, resistance to insects, male fertility, improved nutritional quality, industrial use, stability in the performance and performance improvements.

Duncan et al., Planta, 165: 322-332 (1985), indicate that 97% of the cultivated plants that produced calluses were able to regenerate. In subsequent experiments with inbred and hybrid plants, 91% of regenerable calluses were produced which resulted in plants. In another study, Songstad et al., Plant Cell Reports, 7: 262-265 (1988), described the addition of various media that improved the regenerative capacity of calluses from two inbred lines. In other published reports, "non-traditional" tissues are also indicated to be capable of producing somatic embryogenesis and regeneration of plants. KP Rao et al., Maize Genetics Cooperation Newsletter, 60: 64-65 (1986), relates to somatic embryogenesis of glume callus cultures, and η BV Conger et al., Plant Cell Reports, 6: 345-347 (1987), the somatic embryogenesis of tissue cultures of foliar segments of corn is described. These methods of obtaining plants are frequently used with a high success rate.

Corn tissue culture is described in European Patent Application No. 160390. Methods for cultivating corn tissue that can be adapted for use in rice are also described in Green et al., "Plant Regeneration in Tissue Culture of Maize" , Maize for Biological Research (Plant Molecular Biology Association, Charlottesville, Va., Pp. 367-372, 1982) and in Duncan et al., "The Production of Callus Capable of Plant Regeneration from Immature Embryos of Numerous Zea Mays Genotypes", 165 Plant, 322: 332 (1985). Therefore, another aspect of this invention comprises providing cells that, as they develop and

MEXICAN INSTITUTE 'Jh

OF THE PROPERTY

INDUSTRIAL «“ * - differentiate, produce rice plants that have all or essentially all of the physiological and morphological characteristics of the hybrid rice line “CL261”. See TP Croughan et al., (Springer-Verlag, Berlin, 1991) Rice (Oryza sativa Ly Establishment of Callus Culture and the regeneration of Plants, in Biotechnology in Agriculture and Forestry (19-37).

With the emergence of molecular biology techniques that allow the isolation and characterization of genes encoding specific protein products, it is now possible to modify plants conventionally to incorporate and express foreign genes or additional or modified versions of native or endogenous genes (perhaps targeted by different promoters), in order to alter the characters of a plant in a specific way. These strange, additional and modified genes are collectively referred to herein as transgenes. " Over the past 15-20 years, various methods have been developed to produce transgenic plants, and in particular embodiments, the present invention also relates to transformed versions of "CL261".

An expression vector is constructed that will be able to operate in plant cells. This vector comprises a DNA coding sequence under the control of, or operably linked to, a regulatory element (eg, a promoter). The expression vector may contain combinations of one or more coding sequences / regulatory elements operably linked. Vectors can take the form of a plasmid, and can be used alone or in combination with other plasmids to provide transformed rice plants.

Expression vectors

<img file="MX337901B_D0031.tif" />

Expression vectors commonly include at least one "gene sea operably linked to a regulatory element (eg; —errr promoter), which enables the transformed cells containing the marker to be recovered through negative selection, ie, inhibits growth out of 5 those cells that do not contain the marker gene, or through positive selection, that is, by searching for a product encoded by the genetic marker. Various selection marker genes are commonly known in the art and are commonly used to transform plants, including, for example, genes encoding enzymes that perform metabolic detoxification of a selective chemical inhibitor, such as an antibiotic or herbicide, or genes that encode an altered target that is insensitive to said inhibitor. Positive selection methods are also known in the art.

For example, a selection marker gene that is commonly used to transform plants is that of neomycin phosphotransferase II (nptll), which is isolated from the Tn5 transposon, the expression of which confers resistance to kanamycin. See Fraley et al., Proc. Nati. Acad. Sci. USA, 80: 4803 (1983). Another selection marker gene that is commonly used to transform plants is that of hygromycin phosphotransferase, which confers resistance to the antibiotic hygromycin. See Vanden Elzen et al., Plant Mol. Biol., 5: 299 (1985).

Other selection marker genes of bacterial origin that confer resistance to one or more antibiotics include the genes for gentamicin acetyl transferase, streptomycin phosphotransferase, aminoglycoside-3'-adenyl transferase, and the determinant of bleomycin resistance. Hayford et al., Plant Physiol., 86: 1216 (1988), Jones et al., Mol. Gen. Genet., 210: 86 (1987),

Svab et al., Plant Mol. Biol., 14: 197 (1990), Plant Mol. Biol., 7: 171 (1986). Other gns selection markers confer resistance to pesticides such as

<img file="MX337901B_D0032.tif" />

glyphosate, glufosinate, or bromoxynil. Comai et al., Nature, 317: 741-744 (1985), Gordon-Kamm et al., Plant Cell, 2: 603-618 (1990), and Stalker et al., Science,

242:419-423 (1988).

Selection marker genes for transforming plants that are of non-bacterial origin include, for example, the genes for mouse dihydrofolate reductase, plant 5-enolpiruvilshikimate-3-phosphate synthetase, and plant acetolactate synthetase. Eichholtz et al., Somatic Cell Mol. Genet. 13:67 (1987), Shah et al., Science, 233: 478 (1986), and Charest et al., Plant Cell Rep.,

8:643(1990).

Another class of marker genes to transform plants is based on the search for the supposedly transformed plant cells, instead of a selection based on resistance to a toxic substance such as an antibiotic. These marker genes are particularly useful for quantifying or visualizing the spatial pattern of expression of a gene in specific tissues, and are often referred to as reporter genes, because they can be fused to the desired gene or regulatory sequence. Commonly used Indicator genes include the glucuronidase (GUS), galactosidase, luciferase, chloramphenicol, and acetyltransferase genes. See Jefferson, RA, Plant Mol. Biol. Rep., 5: 387 (1987), Teeri et al., EMBO J., 8: 343 (1989), Koncz et al., Proc. Nati.

Acad. Sci. USA, 84: 131 (1987), and DeBlock et al., EMBO J., 3: 1681 (1984). Another approach to identifying relatively rare transformation events has been the use of a gene encoding a dominant constitutive regulator of the Zea mays anthocyanin pigmentation pathway. Ludwig et al., Science,

247:449 (1990).

The green fluorescent protein (GFP) gene has been used as a marker for gene expression in prokaryotic and ukaryotic cells. See

<img file="MX337901B_D0033.tif" />

Chalfie et al., Science, 263: 802 (1994). GFP and GFP mutants can be used as search markers.

Genes included in expression vectors are driven by a nucleotide sequence that comprises a regulatory element, eg, a promoter. Many appropriate promoters are known in the art, as are other regulatory elements, which can be used alone or in combination with promoters.

As used herein, a "promoter" refers to a region of DNA towards the 5 'end of the transcription start site, which involves recognition and binding of RNA polymerase and other proteins to initiate transcription. A "plant promoter" is a promoter capable of initiating transcription in plant cells. Examples of promoters under developmental control include promoters that initiate transcription preferentially in certain tissues, such as leaves, roots, seeds, fibers, xylem vessels, tracheids, or sclerenchyma. These promoters are known as "tissue preference promoters". Promoters that initiate transcription only in a given tissue are known as "tissue specific promoters." A "promoter with specificity for a cell type" directs expression primarily in certain cell types in one or more organs, for example, vascular cells in the roots or in the leaves. An “inducible” promoter is a promoter that is under the control of the ambient. Examples of environmental conditions that can induce transcription through inducible promoters include anaerobic conditions or the presence of light. Tissue-specific, preferably tissue-specific, cell-type specific and inducible promoters are examples of the "no" promoters.

<img file="MX337901B_D0034.tif" />

constitutive ”. A constitutive promoter is a promoter whose activity under most environmental conditions.

A. Inducible promoters

The inducible promoter is operably linked to the gene to be expressed in rice. Optionally, the inducible promoter is operably linked to a nucleotide sequence encoding a signal sequence, which is operably linked to the gene to be expressed in rice. With an inducible promoter, the transcription rate increases in response to an inducing agent.

It will be possible to use any appropriate inducible promoter in the present invention. See Ward et al., Plant Mol. Biol., 22: 361-366 (1993). Examples include promoters of the ACEI system, which responds to copper, Meft et al., PNAS, 90: 4567-4571 (1993), the In2 gene of maize, which responds to benzenesulfonamide protective herbicides, Hershey et al., Mol. Gen Genetics, 227: 229237 (1991), Gatz et al., Mol. Gen. Genetics, 243: 32-38 (1994), and the repressor T td Tn10, Gatz, Mol. Gen. Genetics, 227: 229-237 (1991). A preferred inducible promoter is one that responds to an inducing agent to which plants normally do not respond, for example, the inducible promoter of a steroid hormone gene, the transcriptional activity of which is induced by a glucocorticosteroid hormone. See Schena et al., Proc. Nati. Acad. Sci., USA 88: 0421 (1991).

B. Constitutive promoters

The constitutive promoter is operably linked to the gene to be expressed by η I rice. Alternatively, the constitutive promoter is attached

<img file="MX337901B_D0035.tif" />

operably to a nucleotide sequence encoding a signal sequence that is operably linked to the gene to be expressed in rice.

Constitutive promoters can also be used in the present invention. Examples include promoters of plant-affecting viruses, such as the cauliflower mosaic virus 35S promoter, Odell et al., Nature, 313: 810-812 (1985), and promoters of the rice actin gene, McEIroy et al., Plant Cell, 2: 163-171 (1990), from ubiquitin, Christensen et al., Plant Mol. Biol., 12: 619-632 (1989), and Christensen et al., Plant Mol. Biol. 18: 675-689 (1992), pEMU, Last et al., Theor. Appl. Genet., 81: 581-588 (1991), MAS, Velten et al., EMBOJ., 3: 2723-2730 (1984), and from histone H3 from maize, Lepetit et al., Mol. Gen. Genetics, 231: 276-285 (1992), and Atanassova et al., Plant Journal, 2 (3): 291-300 (1992).

As the constitutive promoter, the ALS promoter (AHAS) can be used, such as the Xbal / Ncol 5 'fragment of the Brassica napus ALS3 structural gene (or a nucleotide sequence similar to said Xbal / Ncol fragment). See the

PCT application WO 96/30530. The promoter of a rice ALS gene (AHAS) can also be used. See the sequences described in PCT Application WO 01/85970 and in US Patent No. 6943280.

C. Promoters with tissue specificity or tissue preference

The tissue-specific promoter is operably linked to the gene you want to express in rice. Optionally, the tissue specific promoter is operably linked to a nucleotide sequence that encodes a signal sequence that is operably linked to the gene to be expressed in rice. Transformed plants produce the product of transgene expression exclusively in, or in preference to, one or more specific tissues.

<img file="MX337901B_D0036.tif" />

Any promoter with tissue specificity or tissue preference may be used in the present invention. Examples of promoters with tissue specificity or tissue preference include phaseolin gene promoters, Murai et al., Science, 23: 476-482 (1983), and Sengupta-Gopalan et al., Proc. Nati. Acad. Sci. USA, 82: 3320-3324 (1985), a leaf-specific and light-induced promoter, such as that of cab or rubisco, Simpson et al., EMBO J., 4 (11): 2723-2729 ( 1985), and Timko et al., Nature, 318: 579-582 (1985), a promoter with anther specificity, such as that of LAT52, Twell et al., Mol. Gen. Genetics, 217: 240-245 (1989), a promoter with specificity for pollen, such as that of Zm13, Guerrero et al., Mol. Gen. Genetics, 244: 161-168 (1993), or a promoter with a preference for microspores, such as that of apg, Twell et al., Sex. Plant Reprod., 6: 217-224 (1993).

Signal sequences to direct proteins to subcellular compartments

The transport of proteins or peptide molecules produced from the transgenes to a subcellular compartment such as the chloroplast, vacuole, peroxisome, glioxisoma, cell wall or mitochondria, or for secretaries to the apoplast, is carried out by operatively binding the nucleotide sequence encoding the signal sequence to the 5 'or 3' end of the gene encoding the protein or peptide of interest. During protein synthesis and processing, targeting sequences at the 5 'or 3' end of the structural gene can determine the final location of the encoded protein.

Many signal sequences are known in the art. See, for example, Becker et al., Plant Mol. Biol., 20:49 (1992), Cióse, PS, Master's Thesis, Iowa State University (1993), Knox, C. et al., “Structure and Organization of

<img file="MX337901B_D0037.tif" />

Two Divergent Alpha-Amylase Genes from Barley ”, Plant Mol. Biol., 9: 3-17 (1987), Lerner et al., Plant Physiol., 91: 124-129 (1989), Fontes et al., Plant Cell, 3: 483 496 (1991), Matsuoka et al. , Proc. Nati. Acad. Sci., 88: 834 (1991), Gould et al., J. Cell. Biol., 108: 1657 (1989), Creissen et al., Plant J., 2: 129 (1991), Kalderon et al., “A short amino acid sequence able to specify nuclear location”, Cell, 39: 499- 509 (1984), and Steifel et al., "Expression of a maize cell wall hydroxyproline-rich glycoprotein gene in early leaf and root vascular differentiation", Plant Cell, 2: 785793 (1990).

Foreign Protein Genes and Agricultural Genes

Genes of agricultural significance that can be introduced by transformation into rice plants according to the present invention are detailed below.

one. Genes that confer resistance to pests or diseases 15 A. Genes that confer resistance to diseases that affect plants.

Plant defenses are frequently activated by a specific interaction between the product of a disease resistance gene (R) in the plant and the product of a corresponding avirulence gene (Avr) in the pathogen. A plant can be transformed with a cloned resistance gene to design plants resistant to specific strains of a pathogen. See, for example, Jones et al., Science 266: 789 (1994) (cloning the tomato Cf-9 gene to confer resistance to Cladosporium fulvum), Martin et al., Science 262: 1432 (1993) (the Pto gene of the tomato, to confer resistance to Pseudomonas syríngae pv, the tomato encodes a protein kinase), and Mindrinos et al., Cell 78: 1089 (1994) (the RSP2 gene from Arabidopsis, to confer resistance to Pseudomonas syríngae).

<img file="MX337901B_D0038.tif" />

B. A Bacillus thuringiensis protein, a derivative of it, or a synthetic pnlippeptide modeled on the basis of it. See, eg, Geiser et al., Gene 48: 109 (1986), where the cloning and nucleotide sequence of a Bt endotoxin gene is described. DNA molecules encoding endotoxin genes can be obtained in the American Collection of Crop Types, Manassas, Va., for example, under ATCC Accession Numbers 40098, 67136, 31995, and 31998.

C. A lectin. See, eg, Van Damme et al., Plant Molec. Biol. 24:25 (1994), where the nucleotide sequences of various lectin genes that bind to the trick of Olivia miniata are described.

D. A protein that binds to vitamins, such as avidin. See PCT Application US93 / 06487. This publication describes the use of avidin and avidin homologs as larvicides against insect pests.

E. An enzyme inhibitor, eg, a protease or protease inhibitor or an amylase inhibitor. See, eg, Abe et al., J. Biol. Chem. 262: 16793 (1987) (the nucleotide sequence of the rice cysteine ​​proteinase inhibitor), Huub et al., Plant Molec. Biol. 21: 985 (1993) (the nucleotide sequence of a cDNA encoding the tobacco proteinase inhibitor 1), and Sumitani et al., Biosci. Biotech. Biochem. 57: 1243 (1993) (the nucleotide sequence of the Streptomyces nitrosporeus amylase inhibitor).

F. An insect-specific hormone or pheromone, such as ecdysteroid or juvenile hormone, a variant of these, a mimetic based on them, or an antagonist or agonist of these. See, eg, Hammock et al., Natura, 344: 458 (1990), where the expression in baculovirus of a cloned juvenile hormone strain, a juvenile hormone inactivator, is described.

<img file="MX337901B_D0039.tif" />

G. An insect-specific peptide or neuropeptide that, by e&gt; c | =&gt; reserving, alters the physiology of the affected pest. See, for example, Watering^-·, J. Biol. Chem. 269: 9 (1994) (the expression obtained by cloning gives as a res' —lita a DNA that encodes the receptor for the diuretic hormone of insects), Pratt et al., Biochem. Biophys. Res. Comm., 163: 1243 (1989) (an allostatin in E ^ fjDloptera puntata). See also US Patent No. 5266317 to Tomalski et al., Where genes encoding neurotoxins paralyzing rites with specificity for insects are described.

H. A poison with specificity for insects produced in the natural world by a snake, by a wasp, etc. For example, see Pang et af., Gene, 116: 165 (1992), which relates to the heterologous expression in plants of a gene encoding a scorpion peptide that is toxic to insects.

I. An enzyme responsible for the hyperaccumulation of a monoterpane, of a sesquiterpene, of a spheroid, of hydroxamic acid, of a phenylpropanoid derivative or of another non-protein molecule with insecticidal activity.

J. An enzyme that participates in the modification of a molecule with biological activity, including its modification after translation, for example, a glycolytic enzyme, a proteolytic enzyme, a lipolytic enzyme, a r ~ iuicl loop, a cyclase, a transaminase , an esterase, a hydrolase, a phosphatsa, a kinase, a phosphorylase, a polymerase, an elastase, a chitin- &amp; or a glucanase, any of which can be natural or synthetic. XZease PCT Application WO 9302197, Scott et al., Describing the nucleotide sequence of a calase gene. DNA molecules that "contain chitinase-encoding sequences can be obtained, for example, from the American Culture Type Collection under Access numbers 39637 and 67152. See also Kramer et al., Ins ct Biochem. Speckle. Biol. 23: 65 ^ I (1993),

<img file="MX337901B_D0040.tif" />

where the nucleotide sequence of a cDNA encoding tobacco hookworm chitinase is described, and Kawalleck et al., Plant Molec. Biol., 21: 673 (1993), where the nucleotide sequence of the polyubiquitin ub4-2 gene from parsley is described.

K. A molecule that stimulates signal transduction. See, for example, Botella et al., Plant Molec. Biol., 24: 757 (1994), where nucleotide sequences of mango bean calmodulin cDNA clones are described, and Griess et al., Plant Physiol., 104: 1467 (1994), where the sequence of nucleotides of a corn calmodulin cDNA clone.

L. An antimicrobial or unfriendly peptide. See PCT Application WO 9516776 (describing tachiplesin-derived peptides that inhibit plant fungal pathogens) and PCT Application WO 9518855 (describing antimicrobial peptides that confer disease resistance).

M. A membrane permease, a channel former, or a channel blocker. See, eg, Jaynes et al., Plant Sci., 89:43 (1993), where the heterologous expression of a lytic cecropin analog peptide is described to confer resistance to Pseudomonas solanacearum on tobacco plants.

N. An invasive viral protein or a complex toxin derived from it. For example, the accumulation of viral coat proteins in transformed plant cells induces resistance to viral infections or diseases caused by the virus from which the coat protein gene is derived, and also from related viruses. Coated protein mediated resistance has been obtained in plants transformed with proteins from alfalfa mosaic virus, cucumber mosaic virus, tobacco streak virus, potato X virus, potato Y virus, of the ¡_ Μ virus. Ρ · - · £ ...... ··· engraving of tobacco, tobacco necrotic striatum virus and virus in ι · ι | ------ ~~~ ------ '' * · - »---- · πτ tobacco mosaic. See Beachy et ai, Ann. Rev. Phytopathol., 28: 451 (1990).

O. An antibody with insect specificity or an immunotoxin derived from it. In this way, an antibody targeting critical metabolic function in the insect mesenteron inactivates the affected enzyme, thereby eliminating the Insect. See Taylor et al., Abstract No. 497, Seventh International Symposium on Molecular Interactions Between Plants and Microbes (Edinburgh, Scotland, 1994) (an enzyme inactivation in transgenic tobacco mediated by the production of single-chain antibody fragments).

Q. An antibody with specificity for a virus. See, for example, Tavladorakl et al., Nature, 366: 469 (1993), where the protection of transgenic plants where recombinant antibody genes are expressed from virus attacks is described.

Q. A protein that stops development produced in nature by a pathogen or by a parasite. For example, fungal endo-1,4-D-polygalacturonases facilitate fungal colonization and nutrient release by the plant through solubilization of homo-1,4-D-galacturonase in the plant cell wall. See Lamb et ai, Bio / Technology, 10: 1436 (1992). Cloning and characterization of a gene encoding the bean endopolygalacturonase inhibitor protein is described in Toubart et al., Plant J., 2: 367 (1992).

A. A protein that stops development produced in nature by a plant. For example, Logemann et ai, Bio / Technology, 10: 305 (1992), described that transgenic plants where the ribosome inactivating gene of barley is expressed have an increased resistance to fungal diseases.

<img file="MX337901B_D0041.tif" />

2. Genes that confer additional resistance to nn, herhirida ^ beyondL those that are inherent in "CL261", for example, those listed below

A. A herbicide that inhibits growth points or meristems, such as an imidazolinone or a sulfonylurea. Examples of genes in this category encode mutant ALS and AHAS enzymes such as those described, for example, in Lee et al., EMBO J., 7: 1241 (1988), and Miki et al., Theor. Appl. Genet., 80: 449 (1990), respectively. See also US Patent Nos. 5545822, 5736629, 5773703, 5773704, 5952553, 6274796, 6943280,

7019196, 7345221, 7399905 and the International Patent Applications

Published WO 00/27182 and WO 01/85970. Resistance to herbicides acting through it may be due to a different mechanism than a resistant AHAS enzyme. See, for example, US Patent No. 5545822.

B. Glyphosate. Resistance can be conferred by 5-enolpiruvil-315 phosphikimate slntetase (EPSP) and aroA mutant genes. Other phosphono compounds like glufosinate. Resistance can be conferred by phosphinothricin acetyl transferase, PAT, phosphinothricin acetyl transferase genes from Streptomyces hygroscopicus and bar. Pyridinoxy or phenoxy propionic acids and cyclohexones. Resistance can be conferred by genes encoding ACCase inhibitors.

See, for example, US Patent No. 4940835 to Shah et al., Where the nucleotide sequence of a form of EPSP that confers glyphosate resistance is described. It is possible to obtain a DNA molecule encoding a mutant aroA with ATCC Accession Number 39256. The nucleotide sequence of the mutant gene is described in US Patent No. 4769061 to Comai. Sequences are described in European Patent Application No. 0333033 by Kumada et al., And in US Patent No. 4975374 by Goodman et al.

<img file="MX337901B_D0042.tif" />

of glutamine synthetase gene nucleotides that confer resistance to herbicides such as L-phosphinothricin. The nucleotide sequence of a phosphinothricin-acetyl transferase gene is provided in European Application No. 0242246, by Leemans ef al., And in DeGreef et al., Bio / Technology, 7:61 (1989), where described the production of transgenic plants in which chimeric bar genes that encode enzymes with phosphinothricin acetyl transferase activity are expressed. Examples of genes that confer resistance to phenoxy propionic acids and cyclohexones, such as setoxidim and haloxyfop, are the Acc1-S1, Acc1-S2, and Acc1-S3 genes, which are described in Marshall et al., Theor. Appl. Genet.,

83:435(1992).

C. A herbicide that inhibits photosynthesis, such as a triazine (the psbA and gs genes⁺) or a benzonitrile (a nitrilase gene). In Przibilla et al., Plant Cell, 3: 169 (1991), the transformation of Chlamydomonas with plasmids encoding mutant psbA genes is described. Nucleotide sequences of nitrilase genes are described in Stalker US Patent No. 4810648, and the DNA molecules containing these genes are available under ATCC Accession Numbers 53435, 67441 and 67442. Cloning and expression of DNA encoding glutathione S-transferase is described in Hayes et al., Biochem. J.,

285:173(1992).

3. Genes that confer or contribute to a value-added character, for example, those described below

A. A modified fatty acid metabolism, for example, by transforming a plant with a stearyl-ACP desaturase antisense sequence, to increase the plant's stearic acid content. Go

Knultzon et al., Prec. Nati. Acad. Sci. USA 89: 2624 (1992).

B. Lower phytate content

<img file="MX337901B_D0043.tif" />

1) The introduction of a gene that encodes phytase would result in an improvement in the degradation of phytate, which would add more free phosphate to the transformed plant. See, eg, Van Hartingsveldt et al., Gene, 127: 87 (1993), where the nucleotide sequence of an Aspergillus niger phytase gene is described.

2) A gene can be introduced to reduce phytate content. For example, this can be done by cloning and then reintroducing DNA associated with an allele responsible for maize mutants characterized by low levels of phytic acid, or by using a homologous or analogous mutation in rice.

See Raboy et al., Maydica, 35: 383 (1990).

C. The composition of carbohydrates can be modified, for example, by transforming plants with a gene that encodes an enzyme that alters the branching pattern of starch. See Shiroza et al., J. Bacteol., 170: 810 (1988) (a nucleotide sequence of a mutant fructosyltransferase gene from

Streptococcus), Steinmetz et al., Mol. Gen. Genet., 20: 220 (1985) (a nucleotide sequence of a Bacillus subtilis levansacarase gene), Pen et al., Bio / Technology, 10: 292 (1992) (production of transgenic plants where the Bacillus lichenifonnis amylase), Elliot et al., Plant Molec. Biol., 21: 515 (1993) (nucleotide sequences of tomato invertase genes), Sogaard et al., J. Biol. Chem., 268: 22480 (1993) (site-specific mutagenesis of the barley amylase gene I), and Fisher et al., PlantPhysiol., 102: 1045 (1993) (the branching enzyme of corn endosperm starch eleven).

Methods to transform rice

Numerous methods for transforming plants are known in the art, including both biological and physical protocols. See, for example, Miki, et al., “Procedures for Introducing Foreign DNA into Plants”, £ ίΐ Methods fíTPlant Molecular Biology and Biotechnology, Glick BR ​​and Tliuiiipburr, 'J: ~ E; (e ~ d1tores) (CRC Press, Inc., Boca Raton, 1993), pp. 67-88. In addition, expression vectors and methods for performing in vitro culture of plant cells or for transforming and regenerating plant tissue are known in the art. See, for example, Gruber et al., "Vectors for Plant Transformation", in Methods In Plant Molecular Biology and Biotechnology, Glick BR ​​and Thompson, JE (editors) (CRC Press, Inc., Boca Raton, 1993), pp. 89-119.

A. Agrobacterium-mediated transformation

One method of introducing a plant expression vector is based on the Agrobacterium natural transformation system. See, for example, Horsch et al., Science, 227: 1229 (1985). A. tumefaciens and A. rhizogenes are soil bacteria that are pathogenic to plants and that transform plant cells at the genetic level. The Ti and Ri plasmids of A. tumefaciens and A. rhizogenes, respectively, contain the genes responsible for the genetic transformation of plants. See, for example, Kado, CI, Crit. Rev. Plant Sci., 10: 1 (1991). Descriptions of Agrobacterium-based vector systems and methods for performing Agrobacterium-mediated gene transfer are provided in Gruber et al., Supra, Miki et al., Supra, and Moloney, et al., Plant Cell Reports, 8 : 238 (1989). See also the Patent of the

US No. 5591616.

B. Direct gene transfer

Despite the fact that the host range for Agrobacterium-mediated transformation is wide, it is more difficult to transform some 25 species of cultivated cereals and gymnosperms with this gene transfer method, although it has been successful in rice and corn. See

<img file="MX337901B_D0044.tif" />

Hiei et al., The Plant Journal, 6: 271-282 (1994), and US Patent No. 5591616. There are other methods of transforming plants that can be used as alternatives to Agrobacterium-mediated transformation.

One method that can be applied in general to plant transformation is microprojectile-mediated transformation (with what is known as a “gene gun”), where DNA is transported on the surface of microprojectiles, which typically have a diameter of between 1 and 4 pm. The expression vector is introduced into plant tissues with a biolistic device where microprojectiles are accelerated to typical speeds of between 300 and 600 m / s, enough to cross the walls and membranes of plant cells. Sanford et al., Part. Sci. Technol., 5:27 (1987), Sanford, JC, Trends Biotech., 6: 299 (1988), Klein et al., Bio / Technology, 6: 559-563 (1988), Sanford, J.

C., Physiol Plant, 7: 206 (1990), and Klein et al., Biotechnology, 10: 268 (1992). Various target tissues can be bombarded with DNA-coated microprojectiles to produce transgenic plants, including, for example, calluses (type I or type II), immature embryos, and meristematic tissue.

Another method for the physical administration of DNA to plants involves the sonication of the target cells. Zhang et al., Bio / Technology, 9: 996 (1991). As an alternative, a liposome or spheroplast fusion has been used to introduce the expression vectors into the plants. Deshayes et al., EMBO J., 4: 2731 (1985), and Christou et al., Proc Nati. Acad. Sci. USA, 84: 3962 (1987). Direct DNA uptake in protoplasts has also been described using a precipitation with CaCI2, polyvinyl alcohol or poly-Lornitine. Hain et al., Mol. Gen. Genet., 199: 161 (1985), and Draper et al., Plant Cell Physiol., 23: 451 (1982). Electroporation of protoplasts, whole cells, and other tissues has also been described. Donn et al., In the

<img file="MX337901B_D0045.tif" />

Summaries of the Seventh International Congress on Cell and Tissue Culture

Vegetables, IAPTC, A2-38, p. 53 (1990), D'Halluin efaA, ..... PÍañtCeÍL 4: 1495 ^ 15 05 (1992), and Spencer et al., Plant Mol. Biol., 24: 51-61 (1994).

Once the transformation of the rice target tissues has been carried out, by means of the expression of a selection marker gene, the preferential selection of the transformed cells, tissues or plants can be carried out, using regeneration and selection methods known in the art.

These transformation methods can be used to produce an inbred transgenic line. Then, the inbred transgenic line can be crossed with another inbred line (which may have been transformed or may not be transformed) to produce a new inbred transgenic line. As an alternative, it is possible to place a genetic trait that has been entered into one particular rice line on another line, using traditional crossing and backcrossing techniques. For example, backcrossing can be used to place a character entered on a non-elite public line into an elite inbred line, or to put a character from an inbred line that contains an endogenous gene in its genome into one or more inbred lines that are not they contain this gene.

It should be understood that the term "inbred rice plant" also includes single gene conversions from an inbred line. Backcross methods can be used with the present invention to enhance or introduce a characteristic into an inbred line.

Numerous traits based on individual genes have been identified that are not usually subjected to favorable selection in the development of new inbred lines, but which can be improved by crossing and backcrossing. Characters based on individual genes may be transgenic

<img file="MX337901B_D0046.tif" />

or non-transgenic. Examples of these characters include male sterility, waxy starch, herbicide resistance, resistance to diseases of bacterial, fungal or viral origin, insect resistance, male fertility, improved nutritional quality, stability in yield and improved performance. These genes are generally inherited through the nucleus. Known exceptions to nuclear genes include some male sterility genes that are inherited through the cytoplasm, but still functionally act as characters based on individual genes. Various characters based on individual genes are described in US Patent Nos. 5777196, 5948957 and 5969212.

DEPOSIT INFORMATION

On October 8, 2009, a sample of the rice cultivar named “CL261” was deposited in the American Collection of Crop Types (ATCC),

10801 University Boulevard, Manassas, Virginia 20110-2209, and assigned the ATCC Accession number PTA-10388. This deposit was made in accordance with the Budapest Treaty.

OTHER CONSIDERATIONS

Full descriptions of all references cited in this specification are incorporated by reference herein. However, in the event that there is an irreconcilable conflict, this specification will serve as a parameter.