水稻光温敏雄性不育突变体tms3650的鉴定和基因定位

doi:10.16819/j.1001-7216.2023.220109

摘要/Abstract

摘要： 目的雄性不育是水稻杂种优势利用的基础。为进一步解析其调控机制，对一个雄性不育突变体进行了鉴定。方法利用⁶⁰Co-γ对籼稻品种93-11进行辐射诱变，从其后代中鉴定到一个雄性不育突变体tms3650。将籼稻明恢63作父本同突变体tms3650杂交构建F₂和F₃群体，采用图位克隆的方法对目的基因进行精细定位。结果 tms3650突变体其他农艺性状与野生型一致，但花药白绿且瘦小，花粉不能被1%碘-碘化钾溶液染成蓝黑色，穗子包颈，抽穗期延迟。遗传分析表明该突变体表型受一对隐性核基因控制。精细定位结果表明基因位于第3染色体长臂SSR标记RM15927和RM15934之间135.25 kb距离内，且与RM15931标记共分离。陵水冬季南繁鉴定发现，该突变体育性受光温环境影响，说明tms3650突变体的雄性育性在短日低温条件下发生了转育，是一个光温敏雄性不育突变体。结论通过将定位位点与已报道的雄性不育基因比较，发现tms3650是一个新的基因位点，暂命名为TMS3650。

关键词: 水稻, 光温敏雄性不育, 基因定位

Abstract:

【Objective】 Male sterility is the basis of rice heterosis utilization. To further analyze the regulation mechanism, a male sterility mutant was identified.【Method】 The tms3650 mutant was isolated from the mutant library of ⁶⁰Co-γ-treated indica cultivar 93-11. The indica rice Minghui 63 was crossed with the mutant tms3650 as the male parent to construct F₂ and F₃ populations, and the target gene was fine-mapped by map-based cloning. 【Result】 The other agronomic traits of tms3650 were the same as those of the wild type, but the anthers of the mutant were white-green and thin, the pollen could not be stained blue-black by 1% iodine-potassium iodide solution with panicle-enclosure and delayed heading stage. Genetic analysis indicated that the mutant phenotype was controlled by a pair of recessive nuclear genes. The results of fine mapping showed that the gene was located within a distance of 135.25 kb between the SSR markers RM15927 and RM15934 on the long arm of chromosome 3, and co-segregated with the RM15931 marker. The identification of southern breeding in Lingshui in winter revealed that the fertility of mutant was affected by light and temperature, indicating that the tms3650 was a photo-thermo-sensitive male sterile mutant. 【Conclusion】 By comparing with the reported male sterility genes, the tms3650 was controlled by a new gene locus, TMS3650.

Key words: rice, photo-thermo-sensitive male sterility, gene mapping

唐杰, 龙湍, 吴春瑜, 李新鹏, 曾翔, 吴永忠, 黄培劲. 水稻光温敏雄性不育突变体tms3650的鉴定和基因定位[J]. 中国水稻科学, 2023, 37(1): 45-54.

TANG Jie, LONG Tuan, WU Chunyu, LI Xinpeng, ZENG Xiang, WU Yongzhong, HUANG Peijin. Identification and Gene Mapping of a New Photo-thermo-sensitive Male Sterile Mutant tms3650 in Rice[J]. Chinese Journal OF Rice Science, 2023, 37(1): 45-54.

图/表 8

参考文献 59

[1]	朱英国. 水稻雄性不育生物学[M]. 武汉: 武汉大学出版社, 2000.
	Zhu Y G. Biology of Cytoplasmic Male Sterility in Rice[M]. Wuhan: Wuhan University Press, 2000. (in Chinese)
[2]	袁隆平. 水稻的雄性不孕性[J]. 科学通报, 1966(4): 185-188.
	Yuan L P. Male sterility of rice[J]. Science Bulletin, 1966(4): 185-188. (in Chinese with English abstract)
[3]	Darwin C. The Effects of Cross and Self Fertilization in the Vegetable Kingdom[M]. London: John Murray, 1876.
[4]	李晏军.中国杂交水稻技术发展研究(1964-2010)[D]. 南京: 南京农业大学, 2010.
	Li Y J.Research on the Development of Hybrid Rice Technology in China (1964-2010)[D]. Nanjing: Nanjing Agricultural University, 2010. (in Chinese with English abstract)
[5]	石明松. 对光照长度敏感的隐性雄性不育水稻的发现与初步研究[J]. 中国农业科学, 1985(2): 44-48.
	Shi M S. Discovery and preliminary research on recessive male sterile rice sensitive to light length[J]. Scientia Agricultura Sinica, 1985(2): 44-48. (in Chinese with English abstract)
[6]	石明松. 晚粳自然两用系选育及应用初报[J]. 湖北农业科学, 1981(7): 3.
	Shi M S. Preliminary report on breeding and application of late japonica natural dual-purpose line[J]. Hubei Agricultural Science, 1981(7): 3.
[7]	邓华凤, 舒福北, 袁定阳. 安农S-1的研究及其利用概况[J]. 杂交水稻, 1999, 14(3): 1-3.
	Deng H F, Shu F B, Yuan D Y. Research and utilization of Anong S-1[J]. Hybrid Rice, 1999, 14(3): 1-3. (in Chinese)
[8]	杨远柱, 唐平徕, 杨文才, 刘爱民, 陈运泉, 凌文彬, 石天宝. 水稻广亲和温敏不育系株1S的选育及应用[J]. 杂交水稻, 2000, 15(2): 6-7.
	Yang Y Z, Tang P L, Yang W C, Liu A M, Chen Y Q, Ling W B, Shi T B. Breeding and application of 1S of the rice CMS line with broad compatibility and temperature sensitivity[J]. Hybrid Rice, 2000, 15(2): 6-7. (in Chinese with English abstract)
[9]	斯华敏, 付亚萍, 刘文真, 孙宗修, 胡国成. 水稻光温敏雄性核不育系的系谱分析[J]. 作物学报, 2012, 38(3): 394-407.
	Si H M, Fu Y P, Liu W Z, Sun Z X, Hu G C. Pedigree analysis of photoperiod-thermo sensitive genic male sterile rice[J]. Acta Agronimica Sinica, 2012, 38(3): 394-407. (in Chinese with English abstract)
[10]	张华丽, 陈晓阳, 黄建中, 鄂志国, 龚俊义, 舒庆尧. 中国两系杂交水稻光温敏核不育基因的鉴定与演化分析[J]. 中国农业科学, 2015, 48(1): 1-9.
	Zhang H L, Chen X Y, Huang J Z, E Z G, Gong J Y, Shu Q Y. Identification and evolution analysis of photo-thermos sensitive GS gene in Chinese two-line hybrid rice[J]. Scientia Agricultura Sinica, 2015, 48(1): 1-9. (in Chinese with English abstract)
[11]	科技网. 2013年度国家科技奖励:民生类项目成果累累[J]. 科技风, 2013(24): 1.
	Science and Technology Network. 2013 National Science and Technology Awards: People's Livelihood Projects have achieved many achievements[J]. Science and Technology Wind, 2013(24): 1. (in Chinese)
[12]	Chen L, Liu Y G. Male sterility and fertility restoration in crops[J]. Annual Review of Plant Biology, 2014, 65: 579-606. DOI PMID
[13]	Fan Y R, Yang J Y, Mathioni S M, Yu J S, Shen J Q, Yang X F, Wang L, Zhang Q H, Cai Z X, Xu C G, Li X H, Xiao J H, Blake C,. Meyers, Zhang Q F. PMS1T, producing phased small-interfering RNAs, regulates photoperiod-sensitive male sterility in rice[J]. Proceedings of the National Academy of Sciences of the United States of America, 2016, 113(52): 15144-15149. DOI PMID
[14]	梅明华, 陈亮, 章志宏, 李子银, 徐才国, 张启发. 农垦58S光敏不育基因突变位点的确定及pms3区间的进一步作图[J]. 中国科学: C辑, 1999, 29(3): 310-315.
	Mei M H, Chen L, Zhang Z H, Li Z Y, Xu C G, Zhang Q Q. Determination of mutation site of Nongken 58S photosensitive sterility gene and further mapping of pms3 intervals[J]. Science in China: Series C, 1999, 29(3): 310-315. (in Chinese with English abstract)
[15]	Ding J H, Lu Q, Ouyang Y D, Mao H L, Zhang P B, Yao J L, Xu C G, Li X H, Xiao J H, Zhang Q F. A long noncoding RNA regulates photoperiod-sensitive male sterility, an essential component of hybrid rice[J]. Proceedings of the National Academy of Sciences, 2012, 109(7): 2654-2659.
[16]	Zhou H, Liu Q J, Li J, Jiang D G, Zhou L Y, Wu P, Lu S, Li F, Zhu L Y, Liu Z L, Chen L T, Liu Y G, Zhuang C X. Photoperiod- and thermo-sensitive genic male sterility in rice are caused by a point mutation in a novel noncoding RNA that produces a small RNA[J]. Cell Research, 2012, 22(4): 649-660. DOI PMID
[17]	Huang T Y, Wang Z, Hu Y G, Shi S P, Peng T, Chu X D, Shi J, Xiang Z F, Liu D Y. Genetic analysis and primary mapping of pms4, a photoperiod-sensitive genic male sterility gene in rice (Oryza sativa)[J]. Rice Science, 2008(2): 153-156.
[18]	李莉, 邱牡丹, 李懿星, 张大兵, 宋书锋, 王天抗. 一种制备水稻光敏型雄性不育材料的方法及相关基因: CN113637700A[P]. 2021-11-12.
	Li L, Qiu M D, Li Y X, Zhang D B, Song S F, Wang T K. A method for preparing rice photosensitive male sterile material and related genes: CN113637700A[P]. 2021-11-12. (in Chinese)
[19]	Jiang S Y, Cai M, Ramachandran S. Oryza sativa myosin Xi B controls pollen development by photoperiod- sensitive protein localizations[J]. Development Biology, 2007, 304(2): 579 - 592.
[20]	Zhang H, Liang W Q, Yang X J, Luo X, Jiang N, Ma H, Zhang D B. Carbon starved anther encodes a MYB Domain Protein that regulates sugar partitioning required for rice pollen development[J]. The Plant Cell, 2010, 22(3): 672-689. DOI PMID
[21]	李超, 徐建红. 水稻光温敏核不育的分子与表观调控机理[J]. 农业生物技术学报, 2016, 24(1): 115-124.
	Li C, Xu J H. Molecular and epigenetic regulation mechanisms of photothermo-sensitive genic sterility in rice[J]. Journal of Agricultural Biotechnology, 2016, 24(1): 115-124. (in Chinese with English abstract)
[22]	Wang J Z, Wang B, Xu W W. Tagging and mapping rice photoperiod sensitive genic male sterile gene with molecular markers[J]. Plant Genome, 1996, 51(3): 570-577.
[23]	Pitnjam K, Chakhonkaen S, Toojinda T, Muangprom A. Identification of a deletion in tms2 and development of gene-based markers for selection[J]. Planta, 2008, 228(5), 813-822.
[24]	Chutharat C, Ketsuwan C, Keasinee P, Sriprapai C, Numphet S, Kanidta S, Malinee S, Louise V. M, Johnathan A. N, Amorntip M, Rice ORMDL controls sphingolipid homeostasis affecting fertility resulting from abnormal pollen development[J]. PLoS ONE, 2014, 9(9): e106386.
[25]	Subudhi P K, Borkakati R P, Virmani S S, Huang N. Molecular mapping of a thermosensitive genetic male sterility gene in rice using bulked segregant analysis[J]. Genome, 1997, 40(2): 188-194. PMID
[26]	Dong N V, Subudhi P K, Luong P N, Quang V D, Quy T D, Zheng H G, Wang B, H., Nguyen T. Molecular mapping of a rice gene conditioning thermosensitive genic male sterility using AFLP, RFLP and SSR techniques[J]. Theoretical and Applied Genetics, 2000, 100(5): 727-734.
[27]	Reddy O, Siddiq E A, Sarma N P, Ali J, Hussain A J, Nimmakayala P, Ramasamy P, Pammi S, Reddy A S. Genetic analysis of temperature-sensitive male sterility in rice[J]. Theoretical and Applied Genetics, 2000, 100(5): 794-801.
[28]	Wang Y G, Xing Q H, Deng Q Y, Liang F S, Yuan L P, Weng M L, Wang B. Fine mapping of the rice thermo-sensitive genic male-sterile gene tms5[J]. Theoretical and Applied Genetics, 2003, 107(5): 917-921. DOI PMID
[29]	Xu J J, Wang B H, Wu Y H, Du P N, Wang J, Wang M, Yi C D, Gu M H, Liang G H. Fine mapping and candidate gene analysis of ptgms2-1, the photoperiod-thermo-sensitive genic male sterile gene in rice (Oryza sativa L.)[J]. Theoretical and Applied Genetics, 2011, 122(2): 365-372
[30]	Lee D S, Chen L J, Suh H S. Genetic characterization and fine mapping of a novel thermo- sensitive genic male-sterile gene tms6 in rice (Oryza sativa L.)[J]. Theoretical and Applied Genetics, 2005, 111: 1271-1277.
[31]	Wang C H, Zhang P, Ma Z R, Zhang M Y, Sun G C, Ling D H. Development of a genetic marker linked to a new thermo-sensitive male sterile gene in rice(Oryza sativa L.)[J]. Euphytica, 2004, 140: 217-222.
[32]	Liu X, Li X H, Zhang Xin, Wang S W. Genetic analysis and mapping of a thermosensitive genic male sterility gene, tms6(t), in rice(Oryza sativa L.)[J]. Genome, 2010, 53: 119-124.
[33]	Hussain A J, Ali J, Siddiq E A, Gupta V S, Reddy U K, Ranjekar P K. Mapping of tms8 gene for temperature- sensitive genic male sterility (TGMS) in rice (Oryza sativa L.)[J]. Plant Breeding, 2012, 131: 42-47.
[34]	Sheng Z H, Wei X J, Shao G N, Chen M L, Song J, Tang S Q, Luo J, Hu Y C, Hu P S, Chen L Y. Genetic analysis and fine mapping of tms9, a novel thermosensitive genic male-sterile gene in rice (Oryza sativa L.)[J]. Plant Breeding, 2013, 132(2): 159-164.
[35]	Qi Y B, Liu Q L, Zhang L, Mao B Z, Yan D W, Jin Q S, He Z H. Fine mapping and candidate gene analysis of the novel thermo-sensitive genic male sterility tms9-1 gene in rice[J]. Theoretical & Applied Genetics, 2014, 127(5): 1173-1182.
[36]	Li H, Yuan Z, Gema V B, Yang C Y, Liang W Q, Zong J, Wilson Z A, Zhang D B. PERSISTENT TAPETAL CELL1 encodes a PHD-finger protein that is required for tapetal cell death and pollen development in rice[J]. Plant Physiology, 2011, 156(2): 615-630.
[37]	Yu J P, Han J J, Kim Y J, Song M, Yang Z, He Y, Fu R F, Luo Z J, Hu J P, Liang W Q, Zhang D B. Two rice receptor-like kinases maintain male fertility under changing temperatures[J]. Proceedings of the National Academy of Sciences of the United States of America, 2017, 114(46): 12327-12332. DOI PMID
[38]	李月灵. 水稻花粉壁发育相关基因OsACOS12和温敏核不育基因的克隆与功能分析[D]. 上海: 上海师范大学, 2017.
	Li Y L. Cloning and functional analysis of rice pollen wall development-related gene OsACOS12 and thermosensitive sterility gene[D]. Shanghai: Shanghai Normal University, 2017.
[39]	Chang Z Y, Chen Z F, Wang N, Xie G, Lu J W, Yan W, Zhou J L, Tang X Y, Deng X W. Construction of a male sterility system for hybrid rice breeding and seed production using a nuclear male sterility gene[J]. Proceedings of the National Academy of Sciences of the United States of America, 2016, 113(49): 14145-14150. PMID
[40]	李仕贵, 周开达, 朱立煌. 水稻温敏显性核不育基因的遗传分析和分子标记定位[J]. 科学通报, 1999, 44(9): 955-958.
	Li S G, Zhou K D, Zhu L H. Genetic analysis and molecular marker mapping of thermosensitive dominant sterility genes in rice[J]. Science Bulletin, 1999, 44(9): 955-958. (in Chinese with English abstract)
[41]	Koh H J, Son Y H, Heu M H, Lee H S, McCouch S R. Molecular mapping of a new genic male-sterility gene causing chalky endosperm in rice (Oryza sativa L.)[J]. Euphytica, 1999, 106(1): 57-62.
[42]	Jia J H, Zhang D S, Li C Y, Qu X P, Wang S W, Chamarerk V, Nguyen H T, Wang B. Molecular mapping of the reverse thermo-sensitive genic male-sterile gene (rtms1) in rice[J]. Theoretical and Applied Genetics, 2001, 103(4): 607-612.
[43]	Chen R Z, Zhao X, Shao Z, Wei Z, Wang Y Y, Zhu L L, Zhao J, Sun M X, He R F, He G C. Rice UDP-glucose pyrophosphorylase1 is essential for pollen callose deposition and its cosuppression results in a new type of thermosensitive genic male sterility[J]. The Plant Cell, 2007, 19(3): 847-861.
[44]	Zhou Y F, Zhang X Y, Xue Q Z. Fine mapping and candidate gene prediction of the photoperiod and thermo- sensitive genic male sterile gene pmsl(t) in rice[J]. Journal of Zhejiang University, 2011, 12: 436-447.
[45]	Zhou H, Zhou M, Yang Y Z, Li J, Zhu L Y, Jiang D G, Dong J F, Liu Q J, Gu L F, Zhou L Y, Feng M J, Qin P, Hu X C, Song C L, Shi J F, Song X W, Ni E D, Wu X J, Deng Q Y, Liu Z L, Chen M S, Liu Y G, Cao X F, Zhuang C X, RNase ZS1 processes UbL40 mRNAs and controls thermosensitive genic male sterility in rice[J]. Nature Communications, 2014, 5(1): 4884.
[46]	龙湍, 安保光, 李新鹏, 张维, 李京琳, 杨瑶华, 曾翔, 吴永忠, 黄培劲. 籼稻93-11辐射诱变突变体库的创建及其筛选[J]. 中国水稻科学, 2016, 30(1): 44-52. DOI
	Long T, An B G, Li X P, Zhang W, Li J L, Yang Y H, Zeng X, Wu Y Z, Huang P J. Creation and screening of indica rice 93-11 radiation mutant mutant library[J]. Chinese Journal of Rice Science, 2016, 30(1): 44-52. (in Chinese with English abstract)
[47]	Zhao J, Long T, Wang Y F, Tong X H, Tang J, Li J L, Wang H M, Tang L Q, Li Z Y, Shu Y Z, Liu X X, Li S F, Liu H, Li J L, Wu Y Z, Zhang J. RMS2 encoding a gdsl lipase mediates lipid homeostasis in anthers to determine rice male fertility[J]. Plant Physiology, 2020, 182(4): 2047-2064.
[48]	李京琳, 李佳林, 李新鹏, 安保光, 曾翔, 吴永忠, 黄培劲, 龙湍. 水稻ptc1隐性核不育系的创制及其配合力分析[J]. 作物学报, 2021, 47(11): 2173-2183. DOI
	Li J L, Li J L, Li X P, An B G, Zeng X, Wu Y Z, Huang P J, Long T. Creation and combining ability analysis of recessive genic sterile lines with a new ptc1 locus in rice[J]. Acta Agronimica Sinica, 2021, 47(11): 2173-2183. (in Chinese with English abstract)
[49]	李京琳, 李新鹏, 龙湍, 安保光, 曾翔, 吴永忠, 黄培劲. 一个水稻MSP1突变体的鉴定和分析[J]. 植物生理学报, 2018, 54(3): 393-400.
	Li J L, Li X P, Long T, Bao A G, Zeng X, Wu Y Z, Huang P J. Identification and characterization of a rice MSP1 mutant[J]. Journal of Plant Physiology, 2018, 54(3): 393-400. (in Chinese with English abstract)
[50]	Scott O R, Arnold J B. Extraction of DNA from milligram amounts of fresh, herbarium and mummified plant tissues[J]. Plant Molecular Biology, 1985, 5: 69-76. DOI PMID
[51]	Michelmore R. Molecular approaches to manipulation of disease resistance genes[J]. Annual Review of Phytopathology, 1995, 33: 393-427. PMID
[52]	Orjuela J, Garavito A, Bouniol M. A universal core genetic map for rice[J]. Theoretical and Applied Genetics, 2010, 120, 563-572. DOI PMID
[53]	林艳, 刘华清, 付艳萍, 吴明基. 利用TALEN技术编辑水稻光温敏核不育基因PMS3[J]. 福建农业学报, 2019, 34(4): 381-386.
	Lin Y, Liu H Q, Fu Y P, Wu M J. Using TALEN technology to edit rice photothermosensitive sterility gene PMS3[J]. Fujian Agricultural Journal, 2019, 34(4): 381-386. (in Chinese with English abstract)
[54]	王芳权, 范方军, 夏士健, 宗寿余, 郑天清, 王军, 李文奇, 许扬, 陈智慧, 蒋彦婕, 陶亚军, 仲维功, 杨杰. 水稻光温敏核不育基因tms5与pms3的互作效应[J]. 作物学报, 2020, 46(3): 317-329. DOI
	Wang F Q, Fan F J, Xia S J, Zong S Y, Zheng T Q, Wang J, Li W Q, Xu Y, Chen Z Z, Jiang Y J, Tao Y J, Zhong W G, Yang J. Interaction effect of the photothermo-sensitive gene tms5 and pms3 in rice[J]. Acta Agronomica Sinica, 2020, 46(3): 317-329. (in Chinese with English abstract)
[55]	Ding J, Shen J, Mao H, Xie W, Li X, Zhang Q. RNA-directed DNA methylation is involved in regulating photoperiod-sensitive male sterility in rice[J]. Molecular Plant, 2012, 5: 1210-1216. DOI PMID
[56]	Li H J, Kim Y J, Yang L, Liu Z, Zhang J, Shi H T, Huang G Q, Persson S, Zhang D B, Liang W Q. Grass-specific EPAD1 is essential for pollen exine patterning in rice[J]. The Plant Cell, 2020, 32(12): 3961-3977.
[57]	Yang Z F, Sun L P, Zhang P P, Zhang Y X, Yu P, Liu L, Abbas A, Xiang X J, Wu W X, Zhan X D, Cao L Y, Cheng S H. TDR INTERACTING PROTEIN 3, encoding a PHD-finger transcription factor, regulates Ubisch bodies and pollen wall formation in rice[J]. The Plant Journal, 2019, 99(5): 844-861.
[58]	Yu J, Meng Z L, Liang W Q,. Behera S, Kudla J, Matthew R T, Luo Z J, Chen M J, Xu D W, Zhao G C, Wang J, Zhang S Y, Kim Y J, Zhang D B. A rice Ca²⁺ binding protein is required for tapetum function and pollen formation[J]. Plant Physiology, 2016, 172(3): 1772-1786.
[59]	Nonomura K I, Morohoshi A, Nakano M, Eiguchi M, Miyao A, Hirochika H, Kurata N. A germ cell-specific gene of the ARGONAUTE family is essential for the progression of premeiotic mitosis and meiosis during sporogenesis in rice[J]. The Plant Cell, 2007, 19(8): 2583-2594.

育性 Fertility	株系Line									合计 Total
育性 Fertility	L1	L2	L3	L4	L5	L6	L7	L8	L9	合计 Total
可育花粉数Fertile pollen	170	174	156	120	202	71	95	100	39	1127
不育花粉数Sterile pollen	195	215	271	130	190	85	104	81	16	1287
自交收种Selfed seed	240	200	300	320	380	117	194	220	90	2061
结实率Seed setting rate/%	32.0	26.6	40.0	42.6	50.6	15.6	25.8	29.3	12.0	30.5

育性 Fertility	株系Line									合计 Total
育性 Fertility	L1	L2	L3	L4	L5	L6	L7	L8	L9	合计 Total
可育花粉数Fertile pollen	170	174	156	120	202	71	95	100	39	1127
不育花粉数Sterile pollen	195	215	271	130	190	85	104	81	16	1287
自交收种Selfed seed	240	200	300	320	380	117	194	220	90	2061
结实率Seed setting rate/%	32.0	26.6	40.0	42.6	50.6	15.6	25.8	29.3	12.0	30.5

杂交组合 Cross combination	F₁结实率 Seed-setting rate of F₁/%	F₂		χ²_3:1	χ²_0.05
杂交组合 Cross combination	F₁结实率 Seed-setting rate of F₁/%	野生型 Wild type plant	突变体 Mutant plant	χ²_3:1	χ²_0.05
tms3650/93-11	58.13	79	31	0.44	3.841
tms3650/ZH11	76.92	56	14	0.70	3.841
tms3650/明恢63 tms3650/Minghui 63	75.19	186	61	0.01	3.841

杂交组合 Cross combination	F₁结实率 Seed-setting rate of F₁/%	F₂		χ²_3:1	χ²_0.05
杂交组合 Cross combination	F₁结实率 Seed-setting rate of F₁/%	野生型 Wild type plant	突变体 Mutant plant	χ²_3:1	χ²_0.05
tms3650/93-11	58.13	79	31	0.44	3.841
tms3650/ZH11	76.92	56	14	0.70	3.841
tms3650/明恢63 tms3650/Minghui 63	75.19	186	61	0.01	3.841

序号 Serial number	基因 Gene	功能注释 Function annotation
1	LOC_Os03g53570	反转录转座子蛋白 Retrotransposon protein, putative, unclassified, expressed
2	LOC_Os03g53580	表达蛋白 Expressed protein
3	LOC_Os03g53590	表达蛋白 Expressed protein
4	LOC_Os03g53600	HTH DNA结合蛋白 HTH DNA-binding protein, putative, expressed
5	LOC_Os03g53610	晚期胚胎发育丰富的蛋白质 D-34 late embryogenesis abundant protein D-34, putative, expressed
6	LOC_Os03g53620	晚期胚胎发育丰富的蛋白质 D-34 late embryogenesis abundant protein D-34, putative, expressed
7	LOC_Os03g53630	PHD锌指家族蛋白 PHD finger family protein, putative, expressed
8	LOC_Os03g53640	表达蛋白 Expressed protein
9	LOC_Os03g53650	半胱氨酸合酶 Cysteine synthase, putative, expressed
10	LOC_Os03g53660	含有肌球蛋白头部结构域的蛋白质 Myosin head domain containing protein, expressed
11	LOC_Os03g53670	含有YT521-B样家族结构域的蛋白质 YT521-B-like family domain containing protein, expressed
12	LOC_Os03g53680	表达蛋白 Expressed protein
13	LOC_Os03g53690	氧化还原酶，含有短链脱氢酶/还原酶家族结构域的蛋白质 Oxidoreductase, short chain dehydrogenase/reductase family domain containing protein, expressed
14	LOC_Os03g53700	PHD锌指家族蛋白 PHD-finger domain containing protein, putative, expressed
15	LOC_Os03g53710	醛糖1-差向异构酶 Aldose 1-epimerase, putative, expressed
16	LOC_Os03g53720	SRPK4蛋白 SRPK4, putative, expressed
17	LOC_Os03g53730	黄素蛋白wrbA Flavoprotein wrbA, putative, expressed
18	LOC_Os03g53740	表达蛋白 Expressed protein
19	LOC_Os03g53750	核前蛋白A识别因子 Nuclear prelamin A recognition factor, putative, expressed
20	LOC_Os03g53760	核前蛋白A识别因子 Nuclear prelamin A recognition factor, putative, expressed
21	LOC_Os03g53770	含有蛋白质的RNA识别基序 RNA recognition motif containing protein, putative, expressed
22	LOC_Os03g53780	铵转运体2 Ammonium transporter 2, putative, expressed
23	LOC_Os03g53790	周质β-葡萄糖苷酶前体 Periplasmic beta-glucosidase precursor, putative, expressed
24	LOC_Os03g53800	周质β-葡萄糖苷酶前体 Periplasmic beta-glucosidase precursor, putative, expressed