WO2010096975A1 - 一种杂交稻制种方法 - Google Patents

一种杂交稻制种方法 Download PDF

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Publication number
WO2010096975A1
WO2010096975A1 PCT/CN2009/072327 CN2009072327W WO2010096975A1 WO 2010096975 A1 WO2010096975 A1 WO 2010096975A1 CN 2009072327 W CN2009072327 W CN 2009072327W WO 2010096975 A1 WO2010096975 A1 WO 2010096975A1
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Prior art keywords
hybrid rice
grain
sterile line
small
rice
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PCT/CN2009/072327
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English (en)
French (fr)
Inventor
朱旭东
王跃星
倪深
陈红旗
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中国水稻研究所
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Priority claimed from CN2009100964006A external-priority patent/CN101816281B/zh
Priority claimed from CN200910300956A external-priority patent/CN101836580B/zh
Application filed by 中国水稻研究所 filed Critical 中国水稻研究所
Priority to US13/129,540 priority Critical patent/US8889947B2/en
Publication of WO2010096975A1 publication Critical patent/WO2010096975A1/zh

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    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01HNEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H1/00Processes for modifying genotypes ; Plants characterised by associated natural traits
    • A01H1/02Methods or apparatus for hybridisation; Artificial pollination ; Fertility
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01HNEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H5/00Angiosperms, i.e. flowering plants, characterised by their plant parts; Angiosperms characterised otherwise than by their botanic taxonomy
    • A01H5/10Seeds
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01HNEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H6/00Angiosperms, i.e. flowering plants, characterised by their botanic taxonomy
    • A01H6/46Gramineae or Poaceae, e.g. ryegrass, rice, wheat or maize
    • A01H6/4636Oryza sp. [rice]

Definitions

  • the present invention belongs to the field of plant life sciences, and specifically relates to the field of rice breeding technology.
  • hybrids are two genetically stable, genotyped parents, and the seeds produced by sexual crosses (hybrid F1, whose genotypes are completely heterozygous and identical in expression). Plant heterosis is universal, and whether this advantage can be effectively utilized depends to a large extent on whether the cost of producing hybrids and seed production is low, and whether users can purchase.
  • the hybrid rice has been successfully promoted and utilized in China for more than 30 years. In order to obtain higher yield potential of hybrid rice, increasing the grain weight of rice grains is one of the methods used at present, but it also leads to rice farming. The increase in the quantity (weight) increases the cost of seed purchase.
  • hybrid rice Since the successful use of hybrid rice in China in 1974, hybrid rice has been using the method of sowing the parent (recovery) mother (sterile line) and harvesting the parental methods separately. Seeding and harvesting parents, not only the cumbersome operation technology, the cost of labor input, but also the good cross-pollination posture of the father and the mother in the process of seed production, the father is using '(GA 3 ) gibberellin 'The plants are more likely to be lodging than the female parent after treatment, so that the first harvesting of the male parent is not easy to remove the fallen plants or a few fallen tillering ears, resulting in a decrease in the purity of the hybrids (mixing into some restorer lines).
  • hybrid F1 in China is based on the number of days from sowing to heading between the sterile line (mother) and the restorer (parent), and sowing, staging or simultaneous transplanting. Immediately after the end, the father is cut off, or the father is cut off about 25 days after the flowering (to prevent the hybrid from being mixed into the father or even after the flower is thrown), and finally the hybrids on the female parent are mature. Harvest.
  • the production of rice hybrids in this way has been in use for 35 years since 1974. Only seeding, staging, or transplanting parents, staging, or separately harvesting parents will be more labor-intensive than conventional rice production. 4 / acre.
  • a small-grain mutant obtained by r-ray mutagenesis (ZH-sg, 1000-grain weight about 20 g), which is controlled by a single recessive gene sg, the plant height, tillering ability, photosynthetic capacity of sg gene, Spikelet fertility, grain shape, etc. do not produce any adverse collateral effects.
  • the small particle characteristics are controlled by a single recessive gene sg, and the F1 hybrid produced after the mutant material is modified into a sterile line is small.
  • the use of such granules for the improvement of yield potential in conventional rice breeding is not suitable, but the improvement for hybrid rice sterile lines is completely feasible.
  • the object of the present invention is to transfer the small particle mutant gene sg into the sterile line of any form of rice by sexual hybridization, and only to make the kernel of the sterile line smaller, without changing the grain of the rice produced by the hybrid rice. size.
  • the seed production area can be reduced to improve land use efficiency, thereby reducing the cost of rice farmers purchasing and increasing the income of rice farmers.
  • the mixed sowing male parent (recovery) and the female parent (sterile) are used to simplify the hybrid rice seed production process, so that at least the male parent can be removed after the cross-pollination is completed in the rice.
  • the labor of the current separate male parent is reduced by 1-2/mu (that is, compared with the separate male parent, the mixed machinery harvesting per mu can reduce the labor force by 1-2), thereby reducing the seed production.
  • Cost through the reduction of seed production costs to reduce the cost of rice farmers to increase the income of rice farmers. More importantly, it is possible to carry out mechanized production of hybrid rice seed production after mixed harvesting.
  • [10] A method for simplifying hybrid rice seed production procedures to increase seed production efficiency, which in turn comprises the following steps: [11] (1) with rice (, Oryza sativa) ZH-sg (medium flower 11 small mutant) Breeding of small grain sterile line hybrid rice with CGMCC ⁇ 2741 characteristics;
  • Step (1) obtained hybrid rice with small grain sterile lines and large grain, or normal grain type restorer lines Row matching seed production.
  • step (1) of the above-mentioned small-fertilized hybrid rice is based on the hybrid rice containing the small-grain gene sg mutant as a donor parent, and the three-line infertility Maintain hybrid rice, second-line sterile line hybrid rice or other type of sterile line hybrid rice as the recipient parent for hybridization, and obtain the maintainer or sterile line that stably carries the mutant gene sg after 5-7 generations selection.
  • the hybrid rice containing the small-grain gene sg mutant is rice ( Dryza-sativa) ZH-sg (Chinese flower 11 small particle mutant), deposited on CGMCC on November 07, 2008, with the preservation number No. 2741.
  • the small grain sterile line hybrid rice has a thousand grain weight of 19-21 g, or is 25-45% smaller than the 1000-grain weight of the large-grain, normal-type sterile line hybrid rice.
  • the three-line sterile line, the second-line sterile line or other types of sterile line receptors of hybrid rice with normal grain size to large grain size usually have a 1000-grain weight between 25-30 g.
  • the method according to the present invention wherein the three-line male sterile line, the second-type sterile line or other type of sterile line hybrid rice is selected from the group consisting of, for example, sterile line ⁇ -32 ⁇ , Guangzhan 63S, Inner incense 2A, Tianfeng A and conventional varieties / lines P13, NPB and so on.
  • the selection of the 5-7 generations is performed by using molecular marker technology to select the background of the recipient parent by the isolated individual carrying the sg gene, and selecting ⁇ Molecular marker-assisted selection was based on the genotype of the recipient parent.
  • the individuals whose number of grains per panicle exceeds the recipient parent are selected, and the number of grains per panicle of the selected sg gene-bearing maintainer or sterile line is significantly higher than that of the recipient parent to compensate the grain. Miniaturize and balance production levels.
  • the two are sequentially seeded or mixed for the same period, and the seed is advanced.
  • the transplanting restorer line is hybridized with hybrid rice or mixed transplanting.
  • the 26 g recovery system was mixed or seeded.
  • [20] Preferably, according to the method of the invention, it further comprises the steps of: [21] (3) Mixed harvesting, that is, the hybrid F1 matured in the hybrid rice of the small-grain sterile line, mixed harvesting hybrid F1 and restorer grain;
  • the separation may be carried out according to the size of the hybrid F1 and the recovery system, and a suitable mesh sieve may be used.
  • the ZH-sg (Zhonghua 11 small particle mutant) with the accession number CGMCC No. 2741 provided by the present invention was deposited on CGMCC (Ordinary Microbiology Center of China Microbial Culture Collection Management Committee) on November 7, 2008. It is a small particle mutant obtained by r-ray mutagenesis (ZH-sg, 1000-grain weight about 20 g), which is controlled by a single recessive gene sg , plant height, tillering ability, photosynthetic capacity, and small size of rice. Spike fertility, grain shape, etc. do not produce any adverse collateral effects.
  • the F1 hybrid produced by the sterile line hybrid rice is small (because the hybrid is protected by the mother, the sterile line).
  • Genotype control if the male parent is a recovery system with a weight of 26 grams or even more weight, it can be mixed and even live, at least mixed harvesting (parent, female), especially mechanized mixed harvesting, mixed seed after harvesting It can be sorted in commercial seeds, sorted by suitable mesh sieves, and differentiated between hybrids and male parents.
  • hybrids are screened and sieved, because small seeds with a thousand grain weight of less than 20 grams (the female parent) Hybrids) pass through a mesh screen, while normal and large particles cannot pass through the mesh. Since the small particles are controlled by a single recessive gene, the production and expression of hybrid rice production are not affected.
  • This mutational characteristic can be used to introduce the small particle gene sg into any different material by sexual hybridization. If the granule characteristics are introduced into the three-line sterile line hybrid rice, the second-line sterile line hybrid rice or other types of sterile line hybrid rice through hybridization and backcrossing, the operation process of hybrid rice seed production can be simplified to improve hybrid rice. Seed production efficiency. The use of such pellets for the improvement of yield potential in conventional rice breeding is not suitable, but the improvement for hybrid rice sterile lines is completely feasible.
  • the small-grain mutant sg can be used for the improvement of the three-line sterile line (A or B) of nuclear-mass-infertile infertility, and can also be used for environmental interaction-induced light (-temperature) sensitive genic male sterile line (S). And other ways to produce hybrids (such as the 'chemical killing' pathway) in the improvement of sterile lines.
  • the method of the present invention is generally carried out according to the following steps: 1) Breeding of hybrid rice of small-grain sterile line, such as using a small particle mutant ZH-sg of only 19 g of 1000-grain weight as a female parent, and hybridizing with P13 having a particle weight of 25 g.
  • a small granule containing the small-grain mutant gene sg was selected, which was planted into F3' and the subsequent generations were all of the small-grain type, and the basically stable small-grain line and the three-line sterile line hybrid rice such as ⁇ -32 ⁇ , two Hybrid rice, hybrid rice, Guangzhan 63S, etc., crossed and backcrossed to obtain three-line sterile line hybrid rice, second-line sterile line hybrid rice and other types of sterile line hybrid rice containing ZH-sg small particle mutation characteristics; 2) Mixed seeding: seeding with sterile line hybrid rice controlled by a single recessive gene sg, mixed with rice grain normal or large grain, suitable restorer line at maturity, parental or maternal or sequential, or mixed Sowing, or transplanting the male parent in advance, and then broadcasting the female parent later, in short, it can simplify and save labor compared to the current seed production method; 3) Mixed harvesting: After the parents have spent the flowers, they do not need
  • Hybrid rice with small granulocyte gene sg mutant as donor parent hybridized with three-line sterile line hybrid rice, second-line sterile line hybrid rice or other sterile line type hybrid rice receptor parent to obtain hybrid Fl' Planting hybrid Fl', obtaining the F2' generation, the 1000-grain weight of the seeds produced on the contemporary F2' generation plants is maintained as a three-line infertility Hybrid rice, second-line sterile line, hybrid rice or other type of sterile line, horizontal sputum of hybrid rice, 1/4 of individuals isolated in F2' generation carry small grain gene sg , individuals with S g gene are planted into F3' generation Still being a small particle; wherein the hybrid rice containing the small particle gene sg mutant is ZH-sg rice (O ⁇ i- rira), deposited on CGMCC on November 07, 2008, and the preservation number No. 2741;
  • an individual who carries a morphogenetic receptor-like parent carrying a small-grain gene sg backcrossed with the recipient parent or may perform backcrossing as in the A-step procedure, in a backcross BC1F2' generation or a second backcross
  • BC2F2' generation can still isolate 1/4 of small individuals, individuals who have a form like a recipient parent and whose kernels become smaller, that is, individuals carrying the sg gene, can be stably obtained after 5 to 7 generations.
  • Seed cleaning is a necessary process after the production of hybrids in China, with the aim of removing orange stalks, clods, weed seeds and so on. With regard to the production and sale of crop seeds, China has established strict regulations. Generally, hybrid rice seeds are sorted after harvesting, that is, in the form of mesh sieves (main form), impurities such as orange stalks, clods, weed seeds, etc. are taken out. . In the present invention, the separation of the hybrid F1 (small grain) and the male parent (large or normal) can be completed by adding a suitable mesh screen to the cleaning machine. If the current general-purpose hybrid rice seed cleaning machine is used to separate seeds with different grain sizes, the mixed harvesting of the parent and the female parent can greatly simplify the hybrid rice seed production process.
  • the key of the present invention is that the small particle mutation (ZH-sg) is controlled by a single recessive gene, and the contemporary seeds on the F1 plant are controlled by the parental genotype as small particles, while the rice grown by the farmer's hybrid F1 is produced.
  • the grain is controlled by the paternal (recovery) gene, so as long as the paternal control of the grain is normal, the size of the grain formed by F1 is normal.
  • the hybrids of the infertile parents of small granules are contemporary small granules, and the male granules with large or normal granules can be separated by a specific sieve, that is, using the rice seed cleaning machine commonly used in China, the mixed harvested seeds (not Hybrid F1+ restorer from the breeding line is seeded by self-crossing.) After mesh screening, as long as the specific design mesh is installed on the cleaning machine, the hybrid F1 and the male parent can be hybridized with the small grain F1. The self-crossing of the male parent through the mesh and the large grain cannot be completely separated by the mesh.
  • the present invention is mainly applied to the hybrid seed production technology of hybrid rice, and does not have any influence on hybrid rice production.
  • the mutant rice (Oryza-sativa) ZH-sg (Zhonghua 11 small particle mutant) has a 1000-grain weight of 20g (the original parent is 24g/1000 tablets), while the average rice sterile line such as 'Gold 32A' (trade name) has a thousand-grain weight. For 25 g, larger such as ' ⁇ 2A' (trade name) is 30g.
  • the Fl hybrid produced by 1 mu of hybrid rice is 200 kg
  • the number of Fl seeds with a 1000-grain weight of 20 g, 25 g, and 30 g are 10 million, 8 million, and 6.66 million, respectively.
  • rice farmers are generally planting hybrid rice blasts with 1 hole and 1 seedling, 1 mu of field planting seedlings of 15,000 seedlings, and planting 20,000 F1 seeds per acre.
  • the above three thousand-grain weight types of the same weight hybrids can be planted 500 respectively.
  • Mu 400 acres, 333 acres.
  • the above three kinds of 1000-grain weight per mu are converted into weights of 400g, 500g, and 600g, respectively.
  • the seed price (by weight) is 16 yuan/kg
  • the cost is 6.4 yuan, 8.0 yuan, and 9.6 yuan respectively.
  • the mutant ZH-sg (Zhonghua 11 small particle mutant) has a 1000-grain weight of 20 g (the original parent is 24 g/1000 tablets and the restorer of China has a grain weight (1000-grain weight) of 26 g or more.
  • the present invention is only in the existing common cleaning.
  • the addition of a suitable mesh screen on the machine can complete the separation of the hybrid (small grain) and the male parent (large or normal). Separating the seeds with different grain sizes can realize the mixed harvesting of the hybrid rice parent and the female parent. Simplify the hybrid rice seed production process.
  • Figure 1 is a flow chart of Embodiment 1;
  • Figure 3-1 and Figure 3-2 are schematic photographs of the mesh sieves for seed removal and dehydration after seed removal, Figure 3-1 is a round-hole mesh sieve, and Figure 3-2 is a long strip shape. Mesh screen.
  • MCCNo.2741 the date of deposit is November 07, 2008
  • ⁇ -32 ⁇ three-line sterile line, 1000-grain weight 28 g
  • inner incense 2A three lines
  • the sterile line, 30 grams of 1000-grain weight, and 63S second-line sterile line, 1000-grain weight 28 g hybridization, and the improvement of miniaturization of the sterile line.
  • the small particle mutant ZH-sg is used as the donor parent, and II-32A and its corresponding maintainer I
  • I-32B Receptor Parent
  • the corresponding maintainer ⁇ -32 ⁇ of ⁇ -32 ⁇ was crossed with the small particle mutant ZH-sg to obtain the F1' generation.
  • the size of the F1' granule was similar to that of II-32B (normal grain), and F1' was obtained by conventional planting F2'. Generations.
  • 2.1F2' separated 1/4 small-grained individuals and returned to ⁇ -32 ⁇ once to obtain BC1F1' generation.
  • the isolated sg-containing individuals can be subjected to background detection using the molecular marker technique for ⁇ -32 ⁇ genotypes, or by morphological traits to select individuals as similar as ⁇ -32 ⁇ .
  • the characteristics of the new sterile line/maintainer are: 1000-grain weight is 20 g, and the morphology is shown in the upper right side of Figure 2.
  • the selection of the recipient parent may be a three-line maintainer, a secondary sterile line or other forms of sterile hybrid rice, which can be used as a hybrid between the female parent and the donor parent ZH-sg.
  • the grain of the rice planted on the hybrid first generation is similar to the recipient parent.
  • the F2' generation will isolate 1/4 of the individuals carrying the sg gene, and the genes of both donors will be recombined after hybridization. If the improver wants the progeny to be as close as possible to the recipient, it can carry the sg gene.
  • the offspring can be backcrossed 1 to 2 times or 3 to 5 times with the recipient parent, so that the genes of the offspring can be quickly made. Return to the recipient parent.
  • the hybrid rice of the small-germ sterile line and the large- or normal-grained restorer hybrid rice are used for the production of hybrids, 1) if the sterile line (small growth period) and the restorer line (longer growth period) If the growth period differs by 10 days or more, it is still carried out according to the current production mode of hybrid rice seed production (the transplanting restorer is the male parent and the sterile male parent), the male parent (recovery) and the female parent (not Breeding) After the end of the flowering, it is not necessary to separate the male parent separately, but the harvester can be mixed and harvested.
  • the hybrid rice of the small-germ sterile line and the large- or normal-grained restorer hybrid rice are used for the production of hybrids, if the sterile line (small growth period) and restorer line (longer growth period) If the growth period differs within 10 days, the sterile line can be taken without germination, the restorer line is germinated for mixed sowing, or the direct mixed sowing or mixed transplanting.
  • the male parent (recovery system) and the female parent (sterile line) do not have to separate the male parent after the flowering is finished, but can be mixed and harvested with the harvester.
  • the harvested seeds are still carried out according to the current dehydration method of hybrid rice. After the seed water content reaches the specified enthalpy, the mesh with the form of Figure 3-2 is selected, and the appropriate mesh is selected according to the grain width of the sterile line and It is installed on the seed cleaning machine, and the dehydrated standard mixed seed (sterile line and restore line) is put into the seed cleaning machine to separate the hybrid F1 and the male parent (recovery line).
  • the principle is small infertility of the grain. Department (F1) can pass Through the mesh, the large recovery of the grain cannot be separated by the mesh.

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Description

•种杂交稻制种方法 技术领域
[1] 本发明属于植物生命科学领域, 具体涉及水稻育种技术领域。
背景技术
[2] 农作物杂种优势的利用, 必须是以可廉价获得 F1杂交种为前提, 使用者购买杂 交种后种植杂交稻方能获得产量和收益。 所谓杂交种是 2个遗传稳定、 基因型不 同的亲本, 进行有性杂交所产生的种子 (杂交种 F1 , 其基因型完全杂合而表现 性完全一致) 。 植物杂种优势具有普遍性, 这种优势能否被有效利用, 很大程 度上取决于生产杂交种及制种的成本是否低廉, 使用者能否有购买能力。 杂家 水稻在我国成功推广利用已有 30多年, 为求得更高的杂交水稻产量潜力, 提高 稻谷籽粒的粒重是目前被较多釆用的手段之一, 但这也同吋导致了稻农用种量 (重量) 的提升而增加了购种成本。
杂交稻自 1974年在中国成功实现杂种优势利用以来, 一直利用分期播种父 (恢 复系) 母 (不育系) 本并分别收获父母本的方法并沿用至今。 分期播种和收获 父母本, 不仅操作技术繁琐、 增加了劳力投入的成本, 而且在制种过程中为了 父高、 母矮的良好异花授粉姿态, 父本在利用' (GA3) 赤霉素 '处理后植株高于 母本而更易遭致倒伏, 使得先行收割父本吋不易将倒伏的植株或少数倒伏的分 蘖穗清除干净, 从而导致杂交种纯度降低 (混入部分恢复系) 。
[4] 我国目前杂交种 F1的生产均是依据不育系 (母本) 与恢复系 (父本) 之间的播 种至抽穗的历期天数, 进行分期播种、 分期或同期移栽, 待扬花结束后随即割 除父本, 或在扬花后 25天左右先行割除父本 (为防止杂交种混入父本甚至在扬 花后就将父本割除) , 最后待母本上所结的杂交种成熟吋再进行收获。 这一方 式的水稻杂交种生产自 1974年以来已沿用 35年, 仅分期播种、 分期或分别移栽 父母本、 分期或分别收割父母本, 就将在制种田比一般水稻生产多投入劳力 3〜 4个 /亩。
[5] 为提高杂交稻制种效率, 科研人员做了很多研究, 比如中国发明 CN200710070 955公开一种利用双粒突变体提高杂交水稻繁种、 制种效率的方法, 其是通过用 双粒突变体改良不育系实现提高制种效率; CN99101907公开一种杂交水稻制种 方法, 其是通过化学杀雄和除草基因导入的方法来提高制种纯度。 但是, 这些 方法与本申请是根本不同的。
[6] 一种通过 r-射线诱变获得的小粒突变体 (ZH-sg, 千粒重约 20克) , 它受单一隐 性基因 sg控制, sg基因对水稻的植株高度、 分蘖能力、 光合能力、 小穗育性、 籽 粒形状等不产生任何不良的连带影响。 小粒特性受单一隐性基因 sg控制, 用该突 变材料改良成不育系后生产出的 F1杂交种是小粒的。 这种小粒在常规稻育种中 用于产量潜力的改良是不宜的, 但用于杂交稻不育系的改良是完全可行的。 目 前国内外尚无利用小粒隐性突变用于不育系改良的任何报道, 也没有利用籽粒 大小的不同进行混合收割、 并利用种子本身籽粒的大小以网筛的形式进行杂交 种和父本的分拣的报道和发明。
发明内容
[7] 本发明的目的是通过有性杂交, 将小粒突变基因 sg转导入水稻任何形式的不育 系, 仅使不育系自身的籽粒变小, 而不改变杂交稻生产出的稻谷的籽粒大小。 如此, 就可减少制种面积提高土地利用效率, 从而减少稻农购种的成本增加稻 农的收益。 同吋, 在制种过程中实行混合播种父本 (恢复系) 、 母本 (不育系 ) 来简化杂交稻制种程序, 这样至少可以在水稻完成异花授粉后不进行父本的 单独割除, 减少制种过程中由于现行单独割除父本的劳力投入 1-2个 /亩 (即相对 于单独割除父本, 每亩制种田混合机械收割可减少劳力 1-2个) , 从而降低制种 成本, 通过制种成本的降低以减少稻农购种的成本增加稻农的收益。 更为重要 的是, 在实现了混合收割后才有可能进行杂交稻制种的机械化生产。
[8] 本发明是通过下述技术方案实现的:
[9] 扁既 i术
[10] 一种简化杂交稻制种程序提高制种生产效率的方法, 其依次包括下述步骤: [11] ( 1) 具水稻 (、Oryza sativa) ZH-sg (中花 11小粒突变体) CGMCC Νο·2741 特性的小粒不育系杂交稻的育成;
[12] (2) 步骤 (1) 获得的小粒不育系杂交稻与大粒、 或正常粒型恢复系杂交稻进 行配组制种。
[13] 作为优选, 根据本发明所述的方法, 其中, 上述的步骤 (1) 小粒不育系杂交 稻的育成是以含小粒基因 sg突变体杂交稻为供体亲本, 与三系不育保持系杂交稻 、 二系不育系杂交稻或其他类型不育系杂交稻为受体亲本进行杂交, 经 5-7个世 代的选择即可获得稳定携带突变基因 sg的保持系或不育系, 其中, 所述的含小粒 基因 sg突变体杂交稻为水稻 ( Dryza-sativa) ZH-sg (中花 11小粒突变体) , 于 20 08年 11月 07日保藏于 CGMCC , 保藏编号为 No.2741。
[14] 作为优选, 根据本发明所述的方法, 其中, 所述小粒不育系杂交稻其千粒重为 19-21g, 或比大粒、 正常粒型不育系杂交稻的千粒重小 25-45%。 籽粒大小正常 至大粒的杂交稻的三系不育保持系、 二系不育系或其他类型不育系受体亲本其 千粒重通常在 25-30g之间。
[15] 作为优选, 根据本发明所述的方法, 其中, 所述三系不育系、 二系不育系或其 他类型不育系杂交稻选自如不育系 Π-32Α、 广占 63S、 内香 2A、 天丰 A以及常规 品种 /系 P13、 NPB等等。
[16] 作为优选, 根据本发明所述的方法, 其中, 所述的 5-7个世代的选择中, 籍分 子标记技术对分离出的携带 sg基因的个体进行受体亲本的背景选择, 选择吋以受 体亲本的基因型为借鉴进行分子标记辅助选择。 在携带 sg基因的个体选择中, 选 择每穗粒数超过受体亲本的个体, 最终选获的携带 sg基因的保持系或不育系的每 穗粒数明显超过受体亲本, 以补偿籽粒的小型化并平衡产量水平。
作为优选, 根据本发明所述的方法, 其中, 上述步骤 (2) 中的小粒不育系与 恢复系杂交稻的生育期相差小于 10天吋, 二者釆用先后播种或同期混合播种、 先期移栽恢复系杂交稻稍后直播不育系杂交稻或混合移栽。
[18] 作为更优选, 根据本发明所述的方法, 其中, 上述步骤 (2) 中的小粒不育系 与恢复系杂交稻的生育期相差小于 10天吋, 二者釆用同期混合播种、 混合移栽
[19] 作为优选, 根据本发明所述的方法, 其中, 所述的恢复系杂交稻为千粒重大于
26g的恢复系吋釆用混合播种或直播。
[20] 作为优选, 根据本发明所述的方法, 其还包括下述步骤: [21] (3) 混合收割, 即待小粒不育系杂交稻上所结杂交种 F1成熟吋, 混合收割杂 交种 F1与恢复系籽粒;
[22] (4) 种子清选和分离, 即根据杂交种 F1与恢复系籽粒大小不同进行分离。
[23] 作为优选, 根据本发明所述的方法, 其中, 所述根据杂交种 F1与恢复系籽粒大 小不同进行分离釆用适合的网筛即可。
[24] mm
[25] 稻农购买杂交种, 计量单位为公斤 (kg) , 而实质上是满足一定面积种植的杂 交种的颗粒数。 同样是 1000粒种子, 每粒种子的重量不同, 最终总重量是不同 的。 如果杂交种的籽粒是小粒的, 而生产出的杂交稻谷是大粒的 (或正常粒型 的) , 从某种程度上说就可以提高效率和效益。
[26] 本发明提供的保藏编号为 CGMCCNo.2741的 ZH-sg (中花 11小粒突变体) , 于 2 008年 11月 07日保藏于 CGMCC (中国微生物菌种保藏管理委员会普通微生物中 心) , 其是一种通过 r-射线诱变获得的小粒突变体 (ZH-sg, 千粒重约 20克) , 它受单一隐性基因 sg控制, sg基因对水稻的植株高度、 分蘖能力、 光合能力、 小 穗育性、 籽粒形状等不产生任何不良的连带影响。 由于该小粒特性受单一隐性 基因 sg控制, 用该突变材料改良不育系杂交稻后, 此不育系杂交稻生产出的 F1杂 交种是小粒的 (因为杂交种受母体即不育系的基因型控制) , 如果父本是千粒 重在 26克甚至更大粒重的恢复系, 则可以混合播种甚至直播, 至少可以混合收 割 (父、 母本) , 尤其是机械化混合收割, 收割后的混合种子可以在商品种子 进行清选吋通过合适的网筛进行分拣, 区分杂交种和父本, 其原理是杂交种过 网筛清选吋, 因千粒重不足 20克的小粒种子 (母本上结的杂交种) 通过网筛, 而正常粒和大粒无法通过网筛。 由于小粒受单一隐性基因控制, 不影响杂交稻 产量的发挥和表达。
[27] 该突变特性可以通过有性杂交, 将小粒基因 sg导入任何不同的材料中。 若将小 粒特性通过杂交以及回交导入三系不育系杂交稻、 二系不育系杂交稻或其他类 型的不育系杂交稻中, 可以将杂交稻制种的操作过程简化从而提高杂交稻的制 种效率。 这种小粒在常规稻育种中用于产量潜力的改良是不宜的, 但用于杂交 稻不育系的改良是完全可行的。 [28] 为了弥补小粒所带来的产量下降, 在携带 sg基因的个体选择中, 选择每穗粒数 超过受体亲本的个体, 以便最终选获的携带 sg基因的保持系或不育系的每穗粒数 明显超过受体亲本。 禾木科类农作物的产量, 通常情况下由单位面积的有效穗 数、 每穗的实粒数、 以及籽粒的粒重这 3个要素所构成。 3个要素中只要增加某 一个要素的比重, 就有可能使产量增加。 但一般情况下, 增加其中的一个要素 , 会使另外一个或两个要素不同程度的下降, 最终产量的增加极其有限。 因此 , 利用小粒突变体 ZH-sg进行水稻不育系改良, 减少粒重吋必须相应增加每穗粒 数或分蘖数, 即有效穗数。
小粒突变体 sg即可用在核-质互作不育的三系途径不育系 (A或 B) 的改良, 也 可用于环境互作诱导的光 (-温) 敏核不育系 (S) 以及其他方式生产杂交种 (如 '化学杀雄'途径) 的不育系改良中。
[30] 本发明的方法一般按照以下几个步骤进行: 1) 小粒不育系杂交稻育成, 如利 用千粒重仅 19克的小粒突变体 ZH-sg为母本, 与千粒重为 25克的 P13杂交, 在 F2' 中选获了含小粒突变基因 sg的小粒个体, 其种植成 F3'以及后续世代均为小粒类 型, 再将基本稳定的小粒品系与三系不育系杂交稻如 Π-32Α、 二系不育系杂交稻 广占 63S等杂交、 回交, 以获得含 ZH-sg小粒突变特性的三系不育系杂交稻、 二 系不育系杂交稻以及其他类型的不育系杂交稻; 2) 混合播种: 利用受单一隐性 基因 sg控制的不育系杂交稻进行制种, 与稻谷籽粒正常或大粒、 生育期合适的恢 复系配组, 父本与母本或先后、 或同期混合播种, 或先期移栽父本、 稍后直播 母本, 总之可比现在的制种方式简化、 节省劳力; 3) 混合收割: 在父母本扬花 过后无需单独收获父本, 而是用收割机进行混合收割; 4) 网筛分离: 收获后混 合的父本及杂交种 (F1) 在杂交种包装之前的清选工序中, 利用网筛分离千粒 重不同即籽粒大小不同的原理, 可以将不育系上所结的小粒型杂交种与大粒或 籽粒正常的恢复系分离出来。
[31] 具体地说, 上述小粒不育系杂交稻的育成可釆用下述方法:
A) 以含小粒基因 sg突变体杂交稻为供体亲本, 与三系不育保持系杂交稻、 二 系不育系杂交稻或其他不育系类型杂交稻受体亲本杂交, 得到杂种 Fl', 种植杂 种 Fl', 得到 F2'世代, 当代 F2'世代植株上结出的种子千粒重表现为三系不育保持 系杂交稻、 二系不育系杂交稻或其他类型不育系杂交稻水平吋, 在 F2'世代分离 有 1/4的个体携带有小粒基因 sg, 带 Sg基因的个体种植成 F3'世代仍为小粒; 其中 , 所述含小粒基因 sg突变体杂交稻为 ZH-sg水稻 (O ^i- rira) , 于 2008年 11月 07日保藏于 CGMCC, 保藏编号 No.2741;
[33] B) 在 F2'世代选择形态似受体亲本而携带小粒基因 sg的个体, 与受体亲本回交 或可如 A步骤程序执行回交, 在一次回交 BC1F2'世代或二次回交 BC2F2'世代依 然可以分离出 1/4的小粒个体的情况下, 选择形态基本像受体亲本而籽粒变小的 个体, 即携带 sg基因的个体, 经 5〜7个世代的选择即可稳定获得携带突变基因 sg 、 具有小粒特性的不育系或同核保持系杂交稻。
[34] 种子清选是我国目前杂交种生产过后必须的一道工序, 目的是清除桔秆、 土块 、 杂草种子等。 有关农作物种子的生产、 销售, 我国制订了严格的法规, 一般 杂交稻制种收获后要经过清选, 即以网筛的形式 (主要形式) 取出杂质如桔秆 、 土块、 杂草种子等。 本发明只要在清选机上加一道合适的网筛即可完成杂交 种 F1 (小粒) 和父本 (大粒或正常粒) 的分离。 如果用目前的通用型杂交稻种 清选机, 对籽粒大小不同的种子进行分离就能实现混合收获父、 母本, 将极大 简化杂交稻制种程序。
[35] 本发明的关键是小粒突变 (ZH-sg) 受单一隐性基因控制, F1植株上所结当代 种子受母本基因型控制表现为小粒, 而农民种植杂交种 F1所结出的稻谷籽粒受 父本 (恢复系) 基因控制, 故只要父本控制籽粒的基因正常, 其 F1所结籽粒的 大小就正常。 小粒的不育亲本所结杂交种当代为小粒, 与大粒或正常粒型的父 本可以通过特定的筛网进行分离, 即利用目前我国常用的稻谷种子清选机, 对 混合收获的种子 (不育系上所结的杂种 F1+恢复系自交所结的种子) 进行网筛清 选吋, 只要将特定设计网眼安装在清选机上, 杂种 F1和父本自交种子可以因小 粒的杂交种 F1通过网眼、 粒大的父本自交种无法通过网眼而被彻底分离。
[36] 本发明具有如下有益效果:
[37] 本发明主要应用在杂交稻的杂交种制种技术环节, 对杂交稻生产不会造成任何 影响。 突变体水稻 (Oryza-sativa) ZH-sg (中花 11小粒突变体) 的千粒重为 20g (原亲本为 24g/1000粒) , 而一般水稻不育系如 '金 32A' (商品名) 的千粒重为 25 g, 更大的如'内香 2A' (商品名) 为 30g。 假设 1亩杂交稻制种田生产的 Fl杂交种 为 200kg, 千粒重为 20g、 25g、 30g的 Fl代种子数量分别为 1000万粒、 800万粒、 666万粒。 我国稻农在种植杂交稻吋一般是 1穴 1苗, 1亩田栽苗 1.5万棵苗, 以每 亩播种 2万粒 F1种子计, 上述 3种千粒重类型的同等重量杂交种分别能种植 500亩 、 400亩、 333亩。 上述 3种千粒重的每亩用种量按数量折算成重量分别为 400g、 500g、 600g, 当种子价格 (按重量) 同为 16元/ kg吋, 成本分别为 6.4元、 8.0元、 9.6元。
突变体 ZH-sg (中花 11小粒突变体) 的千粒重为 20g (原亲本为 24g/1000粒 而我国的恢复系的籽粒 (千粒重) 都在 26克以上, 本发明只要在现有常用清选 机上加一道合适的网筛即可完成杂交种 (小粒) 和父本 (大粒或正常粒) 的分 离, 对籽粒大小不同的种子进行分离就能实现混合收获杂交稻父、 母本, 将极 大简化杂交稻制种程序。
[39] 如果水稻不育系均釆用携带小粒基 进行制种, 从国家层面上看, 可以节约大 量的制种稻田; 从种业公司层面上看, 可以减少制种过程中的劳动力投入、 简 化操作程序、 减少机械混杂的机会, 同吋还可减少仓储、 运输的数量以及成本 ; 从生产使用者层面上看, 可以减少稻农用于购买杂交稻种子的成本。
附图说明
[40] 图 1是实施例 1流程图;
[41] 图 2是本发明利用小粒突变体改良后的保持系 (图上部, 千粒重 20g, 左为糙米 籽粒, 右为稻谷籽粒。 当与三系的不育系杂交稻杂交进行核置换后即可形成相 应的不育系) ; 以及目前生产上较为普遍的恢复系杂交稻粒型 (图下部, 千粒 重 27g, 左为糙米籽粒, 右为稻谷籽粒) ;
[42] 图 3-1和图 3-2是目前水稻种子繁育收获、 脱水后进行种子清选的网筛示意照片 , 图 3-1为圆孔形网筛、 图 3-2为长条形网筛。
具体实施方式
[43] 下面结合实施例, 更具体地说明本发明的内容。 应当理解, 本发明的实施并不 局限于下面的实施例, 对本发明所做的任何形式上的变通和 /或改变都将落入本 发明保护范围。 [44] 在本发明中, 若非特指, 所有的设备和原料等均可从市场购得或是本行业常用 的。
[45] 实施例 1小粒不育系杂交稻的育成
[46] 以水稻 (Oryza-sativa) ZH-sg (中花 11小粒突变体) 为供体亲本 (保藏编号 CG
MCCNo.2741 , 保藏日期为 2008年 11月 07日) , 可与我国目前杂交稻制种中主要 的不育系 Π-32Α (三系不育系, 千粒重 28克) 、 内香 2A (三系不育系, 千粒重 30 克) 、 广占 63S (二系不育系, 千粒重 28克) 等杂交, 进行不育系籽粒小型化的 改良。
下面结合附图 1, 以小粒突变体 ZH-sg为供体亲本, 以 II-32A及其相应的保持系 I
I-32B (受体亲本) 为例, 说明本发明的实施方式。
1.以 Π-32Α的相应保持系 Π-32Β与小粒突变体 ZH-sg杂交, 得到 F1'世代, F1'粒 型的大小与 II-32B相似 (正常籽粒) , 常规种植 F1'得到 F2'世代。
[49] 2.1F2'分离出 1/4小粒型个体, 并与 Π-32Β回交一次, 得到 BC1F1'世代。 回交前 可对分离出的含 sg个体, 用分子标记技术进行 Π-32Β基因型的背景检测, 或以形 态性状选择与 Π-32Β尽可能相似的个体。
[50] 2.2常规种植 F21导到 F3'世代, 依此类推, 得到 F4'世代、 F5'世代、 F6'世代。
[51] 3.1如果进行一次回交, 则常规种植 BC1F1'得到 BC1F2'世代, 依此类推, 得到 B
C1F3'世代、 得到 BC1F4'世代、 得到 BC1F5'世代。
[52] 3.2可在 BC1F2'中选含 sg基因的个体, 用 Π-32Β再行第 2次回交, 得到 BC2F1'世 代。 在回交吋再次进行背景或行与 Π32Β的相似性选择。
[53] 4 . 常规种植 BC2F1'得到 BC2F2'世代, 依此类推, 得到 BC2F3'世代。
[54] 5.用 II-32A为母本, 与携带 sg基因并农艺形状优异的个体, 即 F4'〜F5'、 BC1F2' 〜BC1F3'、 BC2F2'进行杂交, 即核置换。 以后用携带 sg基因并农艺形状优异的 个体为 F5'〜F6'世代、 BC1F3'〜F4'世代、 BC2F3'世代。 这样的核置换杂交至少 需进行 5次。 从而得到带 sg基因的、 而基因背景与 11-32相似的新型不育系 (A) I 保持系 (B) 。 (说明: 如果想改良的快一些, 可在较低的世代如 F4'就着手与 II
-32A杂交、 回交进行核置换。 本质上没有太大的差别)
新不育系 /保持系的特征为: 千粒重为 20克, 形态如图 2上部右侧所示。 [56] 受体亲本的选择, 可以是三系保持系、 也可以是二系不育系或其他形式的不育 系杂交稻, 均可做母本与供体亲本 ZH-sg杂交, 种植的杂交一代植株上所结稻谷 的籽粒与受体亲本相似。 F2'代会分离出比例占 1/4的携带 sg基因的个体, 因杂交 后受、 供体双方的基因发生重组, 若改良者希望后代尽可能与受体亲本相像, 可以在携带 sg基因的个体中用分子标记技术进行受体基因型背景的选择, 这样可 以提高选择效率。 若希望后代仅保留 sg基因而基因型尽可能与受体亲本相似的话 , 可选带 sg基因的个体与受体亲本回交 1〜2次或 3〜5次, 这样即可迅速使后代的 基因型回归到受体亲本。
[57] 当分离后代携带 sg基因吋, 注意选择穗型比受体亲本大的类型, 这样可使保持 系 (不育系) 在粒重下降后通过增多每穗粒数来弥补, 以平衡或保持原有的产 量潜力。
[58] 实施例 2-1小粒不育系杂交稻的制种
[59] 当小粒不育系杂交稻与大粒或正常粒型的恢复系杂交稻进行杂交种的生产制种 吋, 1) 若不育系 (生育期较短) 与恢复系 (生育期较长) 的生育期相差 10天及 以上的, 仍按目前杂交稻制种的生产方式 (先后分别移栽恢复系父本和不育系 母本) 进行, 父本 (恢复系) 、 母本 (不育系) 扬花结束后不必对父本实行单 独割除, 而可以用收割机混合收割。
[60] 实施例 2-2小粒不育系杂交稻的制种
[61] 当小粒不育系杂交稻与大粒或正常粒型的恢复系杂交稻进行杂交种的生产制种 吋, 若不育系 (生育期较短) 与恢复系 (生育期较长) 的生育期相差在 10天以 内的, 可以釆取不育系不催芽、 恢复系催芽进行混合播种, 或直接混合播种或 混合移栽。 父本 (恢复系) 、 母本 (不育系) 扬花结束后不必对父本实行单独 割除, 而可以用收割机混合收割。
[62] 实施例 3小粒不育系杂交稻的清选与分离
[63] 收获后的种子仍按目前杂交稻的脱水方式进行, 待种子含水量达到规定吋, 选 择具有附图 3-2形式的网筛, 根据不育系的籽粒宽度选择合适的网眼并将其安装 在种子清选机上, 将已脱水达标的混合种子 (不育系与恢复系) 投入种子清选 机, 以分离杂交种 F1和父本 (恢复系) , 其原理是籽粒小的不育系 (F1) 能通 过网眼, 而籽粒大的恢复系无法通过网眼而加以分离。
尽管本发明已经做出了详细的说明并引证了一些具体实施例, 但是对本领域技 术人员来说, 可以对所描述的具体实施例做各种各样的修改和 /或补充或釆取类 似的方式替代是显然的。
Figure imgf000013_0001
下面的说明与本申请说明书中此处提到的
保藏的微生物或其他生物材料相关:
-1 段落号 11
-3 保藏事项
-3-1 保藏单位名称 CGMCC 中国微生物菌种保藏管理委员会普通微生物中心 -3-2 保藏单位地址 中国微生物菌种保藏委员会, 中国北京市 2714信箱, 邮 政编码:100080, Beijing (GN)。
-3-3 保藏日期 2008年 11月 07日 (07.11.2008)
-3-4 保藏号 CGMCC No.2741
-5 本说明是对下列指定国
所有指定国 由受理局填写 -4 本表格与国际申请一起收到:
(是或否)
-4-1 受权官员 由国际局填写 -5 国际局收到本表格日期: -5-1 受权官员

Claims

权利要求书 [1] 一种简化杂交稻制种程序提高制种生产效率的方法, 其特征在于, 所述的 方法依次包括下述步骤:
(1) 具水稻 (O CT ra) ZH-sg (中花 11小粒突变体) CGMCC
No.2741特性的小粒不育系杂交稻的育成;
(2) 步骤 (1) 获得的小粒不育系杂交稻与大粒或正常粒型恢复系杂交稻 进行配组制种。
[2] 如权利要求 1所述的方法, 其特征在于, 所述的步骤 (1) 小粒不育系杂交 稻的育成是以含小粒基因 sg突变体杂交稻为供体亲本, 与三系不育保持系 杂交稻、 二系不育系杂交稻或其他类型不育系杂交稻为受体亲本进行杂交 , 经 5-7个世代的选择即可获得稳定携带突变基因 sg的保持系或不育系, 其 中, 所述的含小粒基因 sg突变体杂交稻为水稻 ZH-sg (中花 11小粒突变体) , 于 2008年 11月 07日保藏于 CGMCC, 保藏编号为 No.2741
[3] 如权利要求 1或 2所述的方法, 其特征在于, 所述小粒不育系杂交稻其千粒 重为 19-21g, 或比大粒、 正常粒型不育系杂交稻的千粒重小 25-45%。
[4] 如权利要求 2所述的方法, 其特征在于, 所述三系不育系、 二系不育系或其 他类型不育系杂交稻选自如不育系 Π-32Α、 广占 63S、 内香 2A、 天丰 A以及 常规品种 /系 P13、 NPB。
[5] 如权利要求 2所述的方法, 其特征在于, 所述的 5-7个世代的选择中, 籍分 子标记技术对分离出的携带 sg基因的个体进行受体亲本的背景选择, 选择 吋以受体亲本的基因型为借鉴进行分子标记辅助选择。
[6] 如权利要求 1所述的方法, 其特征在于, 所述步骤 (2) 中的小粒不育系与 恢复系杂交稻的生育期相差小于 10天吋, 二者釆用先后播种或同期混合播 种、 先期移栽恢复系杂交稻稍后直播不育系杂交稻或混合移栽。
[7] 如权利要求 1或 6所述的方法, 其特征在于, 所述步骤 (2) 中的小粒不育系 与恢复系杂交稻的生育期相差小于 10天吋, 二者釆用同期混合播种、 混合 移栽。
[8] 如权利要求 1所述的方法, 其特征在于, 所述的恢复系杂交稻为千粒重大于
26g的恢复系吋釆用混合播种或直播。
[9] 如权利要求 1所述的方法, 其特征在于, 所述的方法还包括下述步骤:
(3) 混合收割, 即待小粒不育系杂交稻上所结杂交种 F1成熟吋, 混合收割 杂交种 F1与恢复系籽粒;
(4) 种子清选和分离, 即根据杂交种 F1与恢复系籽粒大小不同进行分离。
[10] 如权利要求 9所述的方法, 其特征在于, 所述根据杂交种 F1与恢复系籽粒大 小不同进行分离釆用适合的网筛即可。
PCT/CN2009/072327 2009-02-26 2009-06-18 一种杂交稻制种方法 WO2010096975A1 (zh)

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