WO2018059119A1 - 一种多倍体水稻光温敏雄性核不育系及其选育方法 - Google Patents

一种多倍体水稻光温敏雄性核不育系及其选育方法 Download PDF

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WO2018059119A1
WO2018059119A1 PCT/CN2017/095930 CN2017095930W WO2018059119A1 WO 2018059119 A1 WO2018059119 A1 WO 2018059119A1 CN 2017095930 W CN2017095930 W CN 2017095930W WO 2018059119 A1 WO2018059119 A1 WO 2018059119A1
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rice
tetraploid
male
line
photothermophilic
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French (fr)
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蔡得田
张献华
宋兆建
王维
刘育华
何玉池
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武汉多倍体生物科技有限公司
湖北大学
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Priority to JP2018522521A priority Critical patent/JP2018535672A/ja
Priority to US15/926,156 priority patent/US11051469B2/en
Publication of WO2018059119A1 publication Critical patent/WO2018059119A1/zh

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    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
    • A01H1/02—Methods or apparatus for hybridisation; Artificial pollination ; Fertility
    • A01H1/022—Genic fertility modification, e.g. apomixis
    • A01H1/023—Male sterility
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
    • A01H1/06—Processes for producing mutations, e.g. treatment with chemicals or with radiation
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
    • A01H1/06—Processes for producing mutations, e.g. treatment with chemicals or with radiation
    • A01H1/08—Methods for producing changes in chromosome number
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
    • A01H4/008—Methods for regeneration to complete plants
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H5/00—Angiosperms, i.e. flowering plants, characterised by their plant parts; Angiosperms characterised otherwise than by their botanic taxonomy
    • A01H5/10—Seeds
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H6/00—Angiosperms, i.e. flowering plants, characterised by their botanic taxonomy
    • A01H6/46—Gramineae or Poaceae, e.g. ryegrass, rice, wheat or maize
    • A01H6/4636—Oryza sp. [rice]

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  • the invention belongs to the technical field of breeding new crop varieties in modern agriculture, and particularly relates to a polyploid rice photoperiod sensitive male sterility line and a breeding method thereof.
  • Heterosis is a ubiquitous phenomenon in the biological world.
  • the three-line hybrid rice formed by the hybridization of three lines of rice (male sterile line, male sterile line and male sterile line) completed by scientists represented by Yuan Longping in the 1970s is called the second green revolution.
  • Rice production in China has played a major role in promoting it. Since then, Shi Mingsong (1973) discovered that Hubei light-sensitive genic male sterile line has different manifestations of fertility and infertility in male flower fertility under different lengths of sunshine.
  • This male sterile line in male sterility and short-day low temperature in long-term high temperature is called photo-temperature-sensitive male genic male sterile line; because the sterile line has two characteristics of infertility and fertility, It can be used in two lines, so the hybrid prepared by the sterile line and the restorer line is called a two-line hybrid.
  • the object of the present invention is to provide a method for breeding a polyploid rice photoperiod sensitive male sterile line.
  • Another object of the present invention is to provide a set of polyploid rice photothermophilic male genic male sterile lines.
  • a method for breeding a polyploid rice photothermophilic male genic male sterile line characterized in that the breeding process is:
  • PMeS-1 Sg99012
  • PMeS-2 HN2026
  • PMeS-2 HN2026
  • PMeS-2-2X HN2026-2X
  • PMeS-2-2X HN2026-2X
  • diploid photo-sensitized male genic male sterile line is used as the female parent, and diploid rice with PMeS characteristics is used as the male parent, and diploid hybrid rice is obtained by hybridization;
  • the diploid hybrid rice plants in step b) are doubled into tetraploid hybrid rice; that is, the diploid hybrid rice plants are in the secondary branch differentiation stage of the young panicle differentiation to the meiotic stage.
  • Tissue culture is carried out to form a vigorously growing callus; the callus is transferred to a double culture medium and then differentiated into tetraploid hybrid rice;
  • step d Backcrossing the tetraploid hybrid rice of step c) and the tetraploid photothermophilic male sterility line, and selecting the tetraploid photothermophilic male genic male sterile line from the backcross progeny;
  • the ploidy hybrid rice is emasculated during flowering and backcrossed with the tetraploid photothermophilic male genic male sterile line. Since the rice photothermophilic male sterility is controlled by recessive genes, it still exists in the first generation hybrids after backcrossing. There are fertile plants, so it is necessary to select male plants with good male morphological characteristics according to breeding objectives;
  • the tetraploid photo-temperature-sensitive male genic male sterile rice strain selected in step d) is self-crossed in the fertile period of low-temperature short-day sunshine, and is self-crossed and then hybridized with another tetraploid rice line with PMeS gene.
  • the hybrid hybrid progeny re-selfed, and the tetraploid photo-temperature-sensitive male genic male sterile rice strain was selected; the tetraploid photo-temperature-sensitive male genic male sterile rice plant that was determined in summer was moved to Hainan or artificial climate chamber.
  • the tetraploid photothermophilic male genic male sterile rice strain selected in step e) is continuously selfed for more than one generation in the fertile period of low temperature and short sunshine, at least 6 to 9 generations of continuous selfing;
  • strain having a sterile tetraploid restorative testcross readily available restorer lines for hybrid seed after planting hybrids thereof than the parent, more mothers present heterosis;
  • step g Determine stable and consistent tetraploid photo-temperature-sensitive male genic male sterile rice as a polyploid rice photo-temperature-sensitive male genic male sterile line. After multiple tests and comparisons in step g), the selected morphological characteristics are consistent. Sterile sterility (100% infertility, 99.5% infertility), high seed setting rate (>40%), good heterogeneity, male heterosis with strong heterosis, identified as tetraploid Rice photothermophilic male genic male sterile line, named PSXXX-4x, where XXX is a number.
  • the diploid photothermophilic male genic male sterile line is selected from the following strains: Pei'ai 64S (PA64S), Nongken 58S (Nongken 58S), and HD9802S.
  • the diploid rice having PMeS characteristics is selected from the group consisting of PMeS-1-2X (Sg99012-2X), PMeS-2-2X (HN2026-2X), and PMeS-1-2X (Sg99012-2X). Derived progeny, PMeS-2-2X (HN2026-2X).
  • the tetraploid photothermophilic male sterility line for backcrossing in step d) is obtained by doubling the ear culture of the diploid photothermophilic male genic male sterile line as a male parent A tetraploid photothermophilic male genic male sterile line.
  • the tetraploid rice used for the composite hybridization in the step e) is selected from the following four times the PMeS gene.
  • the tetraploid rice photoperiod sensitive male sterile line selected by the above-mentioned polyploid rice photothermophilic male genic male sterile line has stable morphological characteristics and stable sterility (infertility rate 100) %, infertility 99.5%), high fertility rate (>40%), good heterogeneity, strong heterosis, named PSXXX, where XXX is a number.
  • the rice variety/rice line involved in the breeding method of the present invention is derived from the existing rice varieties/rice lines and their derivatives:
  • Pei'ai 64S (PA64S): Dwarf 64S is a male parent, 58S is used as the female parent, and Pei'ai 64 is used as the male parent.
  • F2 the nuclear sterile plant similar to Pei's 64 is selected and backcrossed with Pei'ai 64.
  • Nongken 58S is a ⁇ -type photoperiod-sensitive genic male sterile line selected by scientist Shi Mingsong in 1973 from the japonica rice variety “Nongken 58”.
  • Huda 9802S Hubei University College of Life Sciences uses "Huda 51" as the female parent, "Hongfu early” as the father's cross, multi-generation breeding and low temperature screening of rice early indica type temperature-sensitive genic male sterility system.
  • PMeS-2(HN2026) The tetraploid rice line PMeS-2 with polyploid meiosis stability (PMeS) selected from the hybrid polyploid progeny of Hubei University of Science and Technology. It has been published in the journal literature "Cai Detian, Chen Jianguo, Chen Dongling et al. Breeding of two polyploid rice lines with polyploid meiosis stability. Chinese Science, 2007, 37(2): 217-226".
  • HN164-4X Hubei University of Science and Technology uses a polyploid rice line PMeS-1 (Sg99012) with polyploid meiosis stability as a parent-derived offspring with meiotic stability and agronomic traits. Polyploid rice line HN164-4X.
  • HN128-4X Polyploid rice line PMeS-1 with polyploid meiosis stability in Hubei University of Science and Technology (Sg99012) is a parental, selected progeny polyploid rice line HN164-4X with meiotic stability characteristics and excellent agronomic traits.
  • A175-4X Hubei University of Science and Technology uses a polyploid rice line PMeS-2 (HN2026) with polyploid meiosis stability as a parent-derived progeny with meiotic stability and good agronomic traits. Polyploid rice line A175-4X.
  • the invention has the beneficial effects that the diploid photothermophilic male genic male sterile line is changed from diploid to tetraploid, and overcomes the low seed setting rate of polyploid rice by crossing with the meiotic stability PMeS strain.
  • the bottleneck problem is to obtain photoperiod sensitive and sterile high-yield tetraploid rice photo-temperature-sensitive male genic male sterile line, which can be used for tetraploid rice hybrid breeding.
  • Figure 2 is a comparison of the flowering of the diploid photothermophilic male genic male sterile line with the tetraploid photothermophilic male sterility line, with the diploid flower on the left and the tetraploid flower on the right.
  • Figure 3 is a pollen fertility of tetraploid rice photothermophilic male genic male sterile line, in which A. infertility period; B. fertility conversion period; C. fertile period.
  • Figure 4 shows tetraploid rice photothermophilic male sterile lines.
  • dF 1 -4X is backcrossed with tetraploid photothermophilic male genic male sterile line HD9802S-4X, that is, using the above-mentioned hybrid plants obtained by doubling, when it is flowering, emasculation and photo-temperature-sensitive male sterility Tetraploid HD9802S-4X (diluted into HD9802S-4X by HD9802S plant) was backcrossed to obtain backcross hybrid BC 1 F 1 ; e. Tetraploid male sterile line was selected from backcross progeny (S strain, Wuhan); f.
  • F 2 is hybridized with another tetraploid rice line HN164-4X (derived offspring of Sg99012) with PMES gene to obtain RCF 1 , and RCF 1 is self-crossed to obtain RCF 2 (abbreviated F 2 ), h.
  • the strain is self-delivered into F 3 , and the male sterile line is self-crossed to obtain F 4 , so that 6 consecutive generations of F 8 are obtained, that is, the selected sterile plants are selected in Wuhan, and the rice indica to Hainan is continuously selfed for 6 generations.
  • anther of tetraploid male sterile line Although it grows much more than the anther of the diploid male sterile line, it is still milky white and arrow-shaped (Fig. 2).
  • Microscopic observation after 0.05% KI-I staining The pollen of the tetraploid sterile plant is round and black in the fertile period, while the pollen is deformed in the infertile period, and the staining is pale yellow or very light grayish black (Fig. 3); the pistil in the flower of the sterile plant is selected.
  • the stigma is more developed, with a unilateral exsertion rate of about 40% and a fertility conversion characteristic with obvious different temperature and light conditions, and the sterility is stable and does not return to the fertile period for more than 25 days; while in the fertile short-day low temperature
  • the seedling has a seed setting rate of 40% or more; in addition, the sterile line and the restored tetraploid restorer line are easy to obtain hybrid seeds (the outcrossing rate is 35% to 45%), and the hybrids have a higher than the parent after planting. The stronger heterosis of the mother; J.
  • the stable tetraploid rice sterile line is a polyploid rice photoperiod sensitive male sterile line, after multiple tests and comparisons of i), breeding Stable morphological characteristics, stable sterility (100% infertility, 99.5% infertility), high seed setting rate (>40%), good heterogeneity, male heterosis with strong heterosis Department, identified as tetraploid rice photothermophilic male genic male sterile line, named PS006, the tetraploid rice PS006 has a sturdy stalk and a dense green leaf color and is uniform (Figure 4).
  • Example 2 The procedure for breeding the tetraploid rice sterile line PS012 is as follows:
  • the tetraploid hybrid F 1 -4X and the photothermophilic male genic male sterile line tetraploid backcross that is, using the above-mentioned hybrid plant obtained by doubling, when it is flowering
  • Male and light-sensitive male genic male sterile line tetraploid PA64S-4X (from PA64S plant seedling culture doubled into PA64S-4X) for backcrossing to obtain backcross first generation hybrid BC 1 F 1 ; e. Select from backcross progeny Tetraploid male sterility sterile line (S strain, Wuhan); f.
  • the strain is self-delivered into F 3 , and the male sterile line is self-crossed to obtain F 4 , so that 6 consecutive generations of F 8 are obtained, that is, the selected sterile plants are selected in Wuhan, and the rice indica to Hainan is continuously selfed for 6 generations.
  • the stable tetraploid rice sterile line is a polyploid rice photoperiod sensitive male sterile line
  • the selected morphological features are stable and stable, and the sterility is stable (100% infertility, 99.5 infertility) )
  • the fertile period seed rate (> 40%), iso with good, strong heterosis with the male sterile line, identified as tetraploid rice light thermo - sensitive genic male sterile line, referred to as PS012.

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Abstract

一种多倍体水稻光温敏雄性核不育系选育方法，所述方法包括：a.确定具有光温敏雄性核不育性和PMeS特性的二倍体水稻品系作为亲本；b.将二倍体光温敏雄性核不育系与二倍体PMeS基因品系杂交，杂种植株幼穗培养加倍成杂种四倍体；c.杂种四倍体与四倍体光温敏雄性核不育系回交；d.从回交后代中选择出四倍体雄性不育株，在低温短日照的可育期自交后再与另一具有PMeS基因的四倍体水稻品系复合杂交；g.挑选四倍体雄性不育株，检测连续自交多代后四倍体雄性不育株的稳定性；i、确定稳定一致的四倍体水稻不育品系为四倍体水稻光温敏雄性核不育系，命名PSXXX。

Description

一种多倍体水稻光温敏雄性核不育系及其选育方法 技术领域
本发明属于现代农业的作物新品种选育技术领域，具体涉及一种多倍体水稻光温敏雄性核不育系及其选育方法。
背景技术
杂种优势是生物界普遍存在的现象。上世纪70年代以袁隆平为代表的科学家完成的水稻三系(雄性不育系、雄性不育保持系和雄性不育恢复系)杂交形成的三系杂交水稻被称为第二次绿色革命，为中国水稻生产起了重大推动作用。此后的石明松(1973)发现的湖北光敏感核不育系，具有在不同日照长度下雄花育性存在可育与不育的不同表现。这种在长日高温下雄性不育、短日低温下雄性可育的不育系，称为光温敏雄性核不育系；由于该不育系具有不育和可育的两种特性，可以一系两用，所以该不育系与恢复系配制的杂种称为两系杂种。
但是，目前全世界应用的水稻都是二倍体，在世界粮食短缺危机形势下，需要大幅度增产，才能解决饥饿问题。而受有性生殖和二倍体性的限制，水稻生产自上世纪80年代以来长期在高产徘徊。为此，蔡得田、袁隆平等(2001)提出“利用远缘杂交和多倍体双重优势选育超级稻”的新战略，确定多倍体水稻亚种间、种间、基因组间杂种优势利用三步战略，把解决多倍体水稻育种结实率低的瓶颈问题作为重中之重的问题来解决。其后，利用组织培养和秋水仙素处理相结合高效率诱导形成多倍体水稻的专利技术和选育出“多倍体减数分裂稳定性PMeS”高结实品系(PMeS多倍体减数分裂稳定性的英文缩写)，突破了多倍体水稻结实率低的瓶颈问题，从而带来多倍体水稻育种的快速发展。大批多倍体水稻材料已经构成，一批多倍体水稻常规品系正进入品种区试阶段。
如果能充分利用多倍体杂种优势，选育多倍体水稻不育系、多倍体水稻恢复系和配制多倍体杂种，将会使水稻生产推向新阶段，对于保障世界粮食安全将具有极其重要的作用。
发明内容
本发明的目的在于提供一种多倍体水稻光温敏雄性核不育系的选育方法。
本发明的另一目的在于提供一组多倍体水稻光温敏雄性核不育系。
为实现上述发明目的，本发明采用的技术方案为：
一种多倍体水稻光温敏雄性核不育系的选育方法，其特征在于，选育过程为：
a.确定具有光温敏雄性核不育性和多倍体减数分裂稳定性PMeS特性的二倍体水稻品系作为亲本；比如典型的光温敏雄性核不育系培矮64S(PA64S)是一种籼型光温敏雄性核不育系，农垦58S(NK58S)是粳型光温敏雄性核不育系，它们具有短日低温条件下可育，而长日高温下不育的育性转换特性，而湖大9802S(HD9802S)是另一种籼型光温敏雄性核不育系，它除了受光照长短影响外，温度起更重要作用，PA64S、HN58S、HD9802S可作为杂交母本；PMeS-1(Sg99012)及其衍生后代、PMeS-2(HN2026)及其衍生后代是具有PMeS特性的四倍体水稻品系，对应的二倍体水稻品系PMeS-1-2X(Sg99012-2X)及其衍生后代、PMeS-2-2X(HN2026-2X)及其衍生后代同样具有多倍体减数分裂稳定性(PMeS)特性，可以作为杂交的父本；
b.以二倍体光温敏雄性核不育系为母本，以具有PMeS特性的二倍体水稻为父本，杂交配制得到二倍体杂种水稻；
c.将b)步骤的二倍体杂种水稻植株幼穗培养加倍成四倍体杂种水稻；即将二倍体杂种水稻植株处于幼穗分化的二次枝梗分化期至减数分裂期的幼穗进行组织培养，形成旺盛生长的愈伤组织；再将愈伤组织转入加倍培养液中培养而后分化成四倍体杂种水稻；
d.将c)步骤的四倍体杂种水稻与四倍体光温敏雄性核不育系回交，从回交后代中选出四倍体光温敏雄性核不育水稻株；即待四倍体杂种水稻开花时去雄，与四倍体光温敏雄性核不育系回交，由于水稻光温敏雄性核不育是隐性基因控制的，因此在回交后一代杂种中还存在有可育植株，因此要根据育种目标挑选雄性不育且形态性状好的水稻植株；
e.将d)步骤选择的四倍体光温敏雄性核不育水稻株在低温短日照的可育期自交，自交后再与另一具PMeS基因的四倍体水稻品系复合杂交，复合杂交后代再自交，从中选出四倍体光温敏雄性核不育水稻株；即将在夏季确定的四倍体光温敏雄性核不育水稻株蔸移至海南或人工气候箱中使其处于低温短日状态恢复育性而自交结实；从这些植株中选择低温短日结实性的植株，使杂种后代不仅含有核不育性基因，而且还含有PMeS基因的高结实性，根据 育种目标挑选雄性不育且且形态性状好的水稻植株；
f.将e)步骤挑选的四倍体光温敏雄性核不育水稻株在低温短日照的可育期连续自交多代，至少连续自交6～9代；
g.检测连续自交多代后的四倍体光温敏雄性核不育水稻株的稳定性，包括染色体数目4X＝48，花药不育的形态特征、花粉育性、柱头性状、光温条件下的育性转换及与四倍体恢复系杂交的异配性和杂种优势；四倍体水稻一般具有茎秆粗壮、叶色浓绿的形态特征，取根尖染色体观察确定4X＝48为四倍体(附图1)；四倍体雄性不育系的花药虽然比二倍体不育系的花药长大许多，但仍然是乳白色、箭头状(附图2)，在0.05％KI-I染色后显微镜观察，四倍体不育株在可育期花粉圆大、黑色，而在不育期则花粉畸形，染色呈浅黄色或极浅的灰黑色(附图3)；挑选不育株的花朵内雌蕊柱头较发达、具有40％左右单边外露率及具明显的不同温光条件下的育性转换特性，而且一旦进入不育期则不育性稳定不返回可育的特点直达25天以上；而在可育的短日低温下具有40％以上的结实率；此外，该不育株与具有恢复性的四倍体恢复系测交容易获得杂交种子、其杂种种植后具有比父、母亲本更强的杂种优势；
h.确定稳定一致的四倍体光温敏雄性核不育水稻为多倍体水稻光温敏雄性核不育系，经过g)步骤的多项检测和比较后，挑选出的形态特征稳定一致、不育性稳定(不育率100％，不育度99.5％)、可育期结实率高(＞40％)、异配性好，杂种优势强的雄性不育系，确定为四倍体水稻光温敏雄性核不育系，命名为PSXXX-4x，其中XXX为数字。
上述方案中，所述二倍体光温敏雄性核不育系选自如下品系：培矮64S(PA64S)、农垦58S(Nongken58S)、HD9802S。
上述方案中，所述具有PMeS特性的二倍体水稻选自如下品系：PMeS-1-2X(Sg99012-2X)、PMeS-2-2X(HN2026-2X)、PMeS-1-2X(Sg99012-2X)的衍生后代、PMeS-2-2X(HN2026-2X)。
上述方案中，步骤d)所述回交用的四倍体光温敏雄性核不育系通过如下方法获得：将作为父本的二倍体光温敏雄性核不育系植株幼穗培养加倍成四倍体光温敏雄性核不育系。
上述方案中，步骤e)所述用于复合杂交的四倍体水稻选自如下具有PMeS基因的四倍 体水稻品系：HN128-4X、HN164-4X、A175-4X。
采用上述多倍体水稻光温敏雄性核不育系的选育方法选育得到的四倍体水稻光温敏雄性核不育系，具有形态特征稳定一致、不育性稳定(不育率100％，不育度99.5％)、可育期结实率高(＞40％)、异配性好、杂种优势强的特性，命名为PSXXX，其中XXX为数字。
本发明所述育种方法中涉及的水稻品种/水稻品系来源于现有已经公开的水稻品种/水稻品系及其衍生品系：
培矮64S(PA64S)：矮64S是以农垦58S作母本、培矮64作父本杂交，在F2选择与培矮64相似的核不育株再与培矮64回交，其杂种后代经长沙、海南多代双向选择育成的籼型水稻低温敏雄性不育系。
农垦58S(NK58S)：农垦58S是我过科学家石明松于1973年从粳稻品种“农垦58”中选育出的粳型光敏核不育系。
湖大9802S(HD9802S)：湖北大学生命科学院用“湖大51”作母本，“红辐早”作父本杂交，经多代选育和低温筛选育成的水稻早籼型温敏核不育系。
PMeS-1(Sg99012)：湖北大学生科院通过几年的籼粳杂交和回交选择与检测，从籼粳杂交多倍体后代中选育出的具有四倍体减数分裂稳定性(polyploid meiosis stability，PMeS)的多倍体水稻品系PMeS-1，已公开于期刊文献“蔡得田，陈建国，陈冬玲等.两个具有多倍体减数分裂稳定性的多倍体水稻品系的选育.中国科学，2007，37(2)：217～226”。
PMeS-2(HN2026)：湖北大学生科院从籼粳杂交多倍体后代中，选育出的具有多倍体减数分裂稳定性(polyploid meiosis stability，PMeS)的四倍体水稻品系PMeS-2，已公开于期刊文献“蔡得田，陈建国，陈冬玲等.两个具有多倍体减数分裂稳定性的多倍体水稻品系的选育.中国科学，2007，37(2)：217～226”。
HN164-4X：湖北大学生科院用具有多倍体减数分裂稳定性的多倍体水稻品系PMeS-1(Sg99012)为亲本选育出的具有减数分裂稳定性特性且农艺性状优良的衍生后代多倍体水稻品系HN164-4X。
HN128-4X：湖北大学生科院用具有多倍体减数分裂稳定性的多倍体水稻品系PMeS-1 (Sg99012)为亲本，选育出的具有减数分裂稳定性特性且农艺性状优良的衍生后代多倍体水稻品系HN164-4X。
A175-4X：湖北大学生科院用具有多倍体减数分裂稳定性的多倍体水稻品系PMeS-2(HN2026)为亲本选育出的具有减数分裂稳定性特性且农艺性状优良的衍生后代多倍体水稻品系A175-4X。
本发明有益效果：本发明将二倍体光温敏雄性核不育系由二倍体变成四倍体，经过与具有减数分裂稳定性PMeS品系杂交而克服多倍体水稻结实率低的瓶颈问题，从而获得光温敏不育和可育期高结实的四倍体水稻光温敏雄性核不育系，可用于四倍体水稻杂种选育。
附图说明
图1为四倍体光温敏雄性核不育系的根尖染色体(2n＝4x＝48)。
图2为二倍体光温敏雄性核不育系与四倍体光温敏雄性核不育系的花器比较，其中左边为二倍体花器，右边为四倍体花器。
图3为四倍体水稻光温敏雄性核不育系花粉育性，其中A.不育期；B.育性转换期；C.可育期。
图4为四倍体水稻光温敏雄性核不育系植株。
具体实施方式
为了更好地理解本发明，下面结合实施例进一步阐明本发明的内容，但本发明的内容不仅仅局限于下面的实施例。
实施例1 四倍体水稻光温敏雄性核不育系PS006选育程序
1.1四倍体水稻不育系PS006选育程序图
Figure PCTCN2017095930-appb-000001
(其中S为光温敏雄性核不育系的简写)
1.2选育程序说明 a.确定具有光温敏雄性核不育性的HD9802S和具有多倍体减数分裂 稳定性(PMeS)的水稻品系HN2026作为亲本；b.二倍体光温敏雄性不育系与二倍体PMeS品系杂交，即利用二倍体不育系HD9802S与HN2026-2X杂交；c.F1杂种植株幼穗培养加倍成杂种四倍体，即采取HD9802S×HN2026-2X杂交的杂种植株处于幼穗分化的二次枝梗分化期至减数分裂期的幼穗进行组织培养，形成旺盛生长的愈伤组织，再将愈伤组织转入加倍培养液中培养而后分化成四倍体杂种F1-4X；d.F1-4X与四倍体光温敏雄性核不育系HD9802S-4X回交，即利用上述已加倍获得的杂种植株，待其开花时去雄与光温敏雄性核不育系四倍体HD9802S-4X(由HD9802S植株幼穗培养加倍成HD9802S-4X)进行回交得到回交一代杂种BC1F1；e.从回交后代中选择出四倍体雄性核不育株(S株，武汉)；f.为确定选择的四倍体雄性核不育株在低温短日照的可育期育性，将在夏季确定的不育株稻蔸移至海南使其处于低温短日状态恢复育性而自交结实，得到BC1F2；g.再将BC1F2与另一具PMeS基因的四倍体水稻品系HN164-4X(Sg99012的衍生后代)复合杂交而得到RCF1，RCF1经自交后得RCF2(简写F2)，h.从中挑选S株自交成F3，雄性不育株系再自交得到F4，如此连续自交6代得到F8，即在武汉进行挑选不育株，带稻蔸至海南繁种连续自交6代得到F8；i.检测F8代四倍体光温敏雄性核不育系的稳定性，包括染色体数目(4X＝48)，花药不育的形态特征、花粉育性、柱头性状，光温条件下的育性转换及与四倍体恢复系杂交的异配性和杂种优势；取根尖染色体观察确定4X＝48为四倍体(附图1)；四倍体雄性不育系的花药虽然比二倍体不育系的花药长大许多，但仍然是乳白色、箭头状(附图2)，在0.05％KI-I染色后显微镜观察，四倍体不育株在可育期花粉圆大、黑色，而在不育期则花粉畸形，染色呈浅黄色或极浅的灰黑色(附图3)；选育不育株的花朵内雌蕊柱头较发达、具有40％左右单边外露率及具明显的不同温光条件下的育性转换特性，而且不育性稳定不返回可育期直达25天以上；而在可育的短日低温下具有40％以上的结实率；此外，该不育株与具有恢复性的四倍体恢复系测交容易获得杂交种子(异交率35％～45％)、其杂种种植后具有比父、母亲本更强的杂种优势；J.确定该稳定一致的四倍体水稻不育品系为多倍体水稻光温敏雄性核不育系，经过i)的多项检测和比较后，选育出的形态特征稳定一致、不育性稳定(不育率100％，不育度99.5％)、可育期结实率高(＞40％)、异配性好，所配杂种优势强的雄性不育系，确 定为四倍体水稻光温敏雄性核不育系，命名为PS006，该四倍体水稻PS006具有茎秆粗壮、叶色浓绿的形态特征且整齐一致(附图4)。
实施例2 四倍体水稻不育系PS012选育程序说明如下：
1.1四倍体水稻不育系PS012选育程序图
Figure PCTCN2017095930-appb-000002
(其中S为光温敏雄性核不育系的简写)
1.2选育程序说明 a.确定具有光温敏雄性核不育性的PA64S和具有多倍体减数分裂稳定性(PMeS)的水稻品系HN2026-2X作为亲本；b.二倍体光温敏雄性不育系与二倍体PMeS品系杂交，即利用二倍体不育系PA64S与HN2026-2X杂交；c.F1杂种植株幼穗培养加倍成杂种四倍体F1-4X，即采取PA64S×HN2026-2X杂交的杂种植株处于幼穗分化的二次枝梗分化期至减数分裂期的幼穗进行组织培养，形成旺盛生长的愈伤组织，再将愈伤组织转入加倍培养液中培养而后分化成四倍体杂种F1-4X；d.四倍体杂种F1-4X与光温敏雄性核不育系四倍体回交，即利用上述已加倍获得的杂种植株，待其开花时去雄与光温敏雄性核不育系四倍体PA64S-4X(由PA64S植株幼穗培养加倍成PA64S-4X)进行回交得到回交一代杂种BC1F1；e.从回交后代中选择出四倍体雄性核不育株(S株，武汉)；f.为确定选择的四倍体雄性核不育株在低温短日照的可育期育性，将在夏季确定的不育株稻蔸移至海南使其处于低温短日状态恢复育性而自交结实，得到BC1F2；g.将BC1F2再与另一具PMeS基因的四倍体水稻品系A175-4X(HN2026的衍生后代)复合杂交而得到RCF1，RCF1经自交后得RCF2(简写F2)，h.从中挑选S株自交成F3，雄性不育株系再自交得到F4，如此连续自交6代得到F8，即在武汉进行挑选不育株，带稻蔸至海南繁种连续自交6代得到F8；i.检测F8代四倍体光温敏雄性核不育系的稳定性，包括染色体数目(4X＝48)，花药不育的形态特征、花粉育性、柱头性状，光温条件下的育性转换及与四倍体恢复系杂交的异配性和杂种优势；J.确定该稳定一致的四倍体水稻不育品系为多倍体水稻光温敏雄性核不育系，经过i)的多项检测和比较后， 选育出的形态特征稳定一致、不育性稳定(不育率100％，不育度99.5％)、可育期结实率高(＞40％)、异配性好，所配杂种优势强的雄性不育系，确定为四倍体水稻光温敏雄性核不育系，简称为PS012。
显然，上述实施例仅仅是为清楚地说明所作的实例，而并非对实施方式的限制。对于所属领域的普通技术人员来说，在上述说明的基础上还可以做出其它不同形式的变化或变动。这里无需也无法对所有的实施方式予以穷举。而因此所引申的显而易见的变化或变动仍处于本发明创造的保护范围之内。

Claims (7)

  1. 一种多倍体水稻光温敏雄性核不育系的选育方法，其特征在于，选育过程为：
    a.确定具有光温敏雄性核不育性和具有PMeS特性的二倍体水稻品系作为亲本；
    b.以二倍体光温敏雄性核不育系为母本，以具有PMeS特性的二倍体水稻为父本，杂交配制得到二倍体杂种水稻；
    c.将b)步骤的二倍体杂种水稻植株幼穗培养加倍成四倍体杂种水稻；
    d.将c)步骤的四倍体杂种水稻与四倍体光温敏雄性核不育系回交，从回交后代中选育出四倍体光温敏雄性核不育水稻；
    e.将d)步骤选择的四倍体光温敏雄性核不育水稻在低温短日照的可育期自交，自交后代再与另一具有PMeS基因的四倍体水稻品系复合杂交，复合杂交后代再自交，从中选出四倍体光温敏雄性核不育水稻；
    f.将e)步骤挑选的四倍体光温敏雄性核不育水稻株在低温短日照的可育期连续自交多代；
    g.检测f)步骤中连续自交多代后的四倍体光温敏雄性核不育水稻株的稳定性，包括染色体数目4X＝48，花药不育的形态特征、花粉育性、柱头性状、光温条件下的育性转换及与四倍体恢复系杂交的异配性和杂种优势；
    h.确定稳定一致的四倍体光温敏雄性核不育水稻为多倍体水稻光温敏雄性核不育系，命名为PSXXX，其中XXX为数字。
  2. 根据权利要求1所述多倍体水稻光温敏雄性核不育系的选育方法，其特征在于，所述二倍体光温敏雄性核不育系选自如下品系：培矮64S、农垦58S、HD9802S。
  3. 根据权利要求1所述多倍体水稻光温敏雄性核不育系的选育方法，其特征在于，所述具有PMeS特性的二倍体水稻选自如下品系：PMeS-1-2X(Sg99012-2X)、PMeS-1-2X(Sg99012-2X)的衍生后代、PMeS-2-2X(HN2026-2X)、PMeS-2-2X(HN2026-2X)的衍生后代。
  4. 根据权利要求1所述多倍体水稻光温敏雄性核不育系的选育方法，其特征在于，步骤d)所述回交用的四倍体光温敏雄性核不育系通过如下方法获得：将作为母本的二倍体光温敏雄性核不育系植株幼穗培养加倍成四倍体光温敏雄性核不育系。
  5. 根据权利要求1所述多倍体水稻光温敏雄性核不育系的选育方法，其特征在于，步骤e)中所述用于复合杂交的四倍体水稻选自如下具有PMeS基因的四倍体水稻品系：HN128-4X、HN164-4X、A175-4X。
  6. 根据权利要求1所述多倍体水稻光温敏雄性核不育系的选育方法，其特征在于，步骤f)中所述连续自交多代是指连续自交至少6～9代。
  7. 权利要求1～6任一所述多倍体水稻光温敏雄性核不育系的选育方法选育得到的四倍体水稻光温敏雄性核不育系，具有形态特征稳定一致、不育性稳定、可育期结实率高、异配性好、杂种优势强的特性，命名为PSXXX，其中XXX为数字。
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