Mechanism of Sterility and Breeding Strategies for Photoperiod/Thermo-Sensitive Genic Male Sterile Rice

To understand the male sterility mechanism of photoperiod/thermo-sensitive genic male sterile [P（T）GMS] lines in rice, the research progress on genetics of photoperiod and/or temperature sensitive genic male sterility in rice was reviewed. A new idea was proposed to explain the sterility mechanism of P（T）GMS rice. The fertility transition from sterile to fertile is the result of cooperative regulation of major-effect sterile genes with photoperiod and/or temperature sensitive genes, but not the so-called pgms gene in P（T）GMS rice. The minor-effect genes, which exhibit accumulative effect on sterility, are the important factors for the critical temperature of sterility transition. The more minor-effect genes the sterile line holds, the lower the critical temperature of sterility transition is. The critical temperature of sterility transition will be invariable if all the minor-effect genes are homozygous. The strategies for breeding P（T）GMS rice were also proposed. The selective indices of critical photoperiod and temperature for sterility transition should be set according to varietal type and ecological region. Imposing selection pressure is a key technology for breeding P（T）GMS rice with lower critical temperature for sterility, and improving the comprehensive performance of the whole traits and combining ability is vital for breeding P（T）GMS rice lines.

Plant Temperature and Its Simulation Model of Thermo-Sensitive Genic Male Sterile Rice

Plant temperature （Tp） and its relations to the microclimate of rice colony and irrigation water were studied using a thermo-sensitive genic male sterile （TGMS） rice line, Pei＇ai 64S. Significant differences in the daily change of temperature were detected between Tp and air temperature at the height of 150 cm （TA）. From 8：00 to 20：00, Tp was lower than TA, but they were similar during 21：00 to next 7：00. The maximum Tp occurred one hour earlier than the maximum TA, though they both reached the minimum at 6：00. Tp fluctuated less than TA. At the same height, during 6：00-13：00, Tp was higher than air temperature （Ta）, and Tp reached the maximum one hour earlier than Ta. During the rest time on sunny day, Tp was close to or even a little lower than Ta. On overcast day, Tp was higher than Ta in the whole day, and both maximized at the same time. In addition, Tp was regulated by solar radiation, cloudage and wind speed in daytime, and by irrigation water at night. The present study indicated that a TA of 29.6℃ was the critical point, at which Tp was increased or decreased by irrigation water. Tp and the difference between water and air temperatures showed a conic relation. Tp fluctuation was also regulated by the absorption or reflection of solar radiation by leaves during daytime and release of heat energy during nighttime. By analysis on correlation and regression simulation, two models of Tp were established.

New Temperature Sensitive Genic Male Sterile Lines with Better Outcrossing Ability for Production of Two-Line Hybrid Rice

An investigation was carried out with three newly developed temperature sensitive genic male sterile （TGMS） lines for their floral traits, seed production potential and outcrossing ability in ten cross combinations. In the TGMS lines, fertile pollens had an average diameter of 0.89 mm while the sterile pollens was with 0.02 mm diameter.TS-29-150GY produced the biggest fertile pollens with 0.92 mm and other two lines produced relatively smaller pollens with 0.91 and 0.85 ram. Pollen fertility during the fertility reversion period was an average of 60.7%. TS-29-150GY had the maximum of 66.9% spikelet fertility whereas other two lines （TNAU18S and TNAU60S） had relatively lower spikelet fertility of 27.8% and 26.7%, respectively. Average of 17.00 g of seed yield was obtained in the TGMS lines during the fertility reversion period. TS-29- 150GY had the highest value of 21.20 g of seed yield while TNAU18S and TNAU60S produced 16.6 g and 13.2 g of seed yield, respectively. The low seed production ability of these three TGMS lines was attributed only to the environmental conditions prevailing during the period. All three TGMS lines had considerable outcrossing potential of 41.2%, 24.6% and 25.0%, respectively. The cross combinations viz. TNAU18S/IET21508 （36 g/plant）, TNAU18S/IET21044 （13 g/plant）, TNAU18S/IET21009 （26.5 g/plant）, TNAU60S/CB-09-106 （26.2 g/plant）, TNAU60S/IET21009 （14 g/plant） and TS29-150-GY/DRR 3306 （39.2 g/plant） showed perfect synchronization with acceptable hybrid seed yield, indicating suitability of TGMS system under Indian condition. Based on the outcrossing related traits viz. panicle exertion, angle of glume opening, stigma length and pollen size, TNAU18S was identified as the best, followed by TS-29-150GY.

Molecular Markers and Candidate Genes for Thermo-Sensitive Genic Male Sterile in Rice

The discovery of thermo-sensitive genic male sterility(TGMS) has led to development of a simple and highly efficient two-line breeding system. In this study, genetic analysis was conducted using three F_2 populations derived from crosses between IR68301 S, an indica TGMS rice line, and IR14632(tropical japonica), Supanburi 91062(indica) and IR67966-188-2-2-1(tropical japonica), respectively.Approximately 1:3 ratio between sterile and normal pollen of F_2 plants from the three populations revealed that TGMS is controlled by a single recessive gene. Bulked segregant analysis using simple sequence repeat(SSR) and insertion-deletion(InDel) markers were used to identify markers linked to the tms gene. The linkage analysis based on the three populations indicated that the tms locus was located on chromosome 2 covering the same area. Using IR68301S × IR14632 F_2 population, the results showed that the tms locus was located between SSR marker RM12676 and InDel marker 2gAP0050058. The genetic distance from the tms gene to these two flanking markers were 1.10 and 0.82 cM, respectively.InDel marker 2gAP004045 located between these two markers showed complete co-segregation with the TGMS phenotype. In addition, InDel marker vf0206114052 showed 2.94 cM linked to the tms gene using F_2 populations of IR68301S × Supanburi 91062. These markers are useful tool for developing new TGMS lines by marker-assisted selection. There were ten genes located between the two flanking markers RM12676 and 2gAP0050058. Using quantitative real-time PCR for expression analysis, 7 of the 10 genes showed expression in panicles, and response to temperatures. These genes could be the candidate gene controlling TGMS in IR68301S.

Improving Blast Resistance of a Thermo-Sensitive Genic Male Sterile Rice Line GD-8S by Molecular Marker-Assisted Selection

The broad-spectrum blast resistance gene Pi-1, from donor line BL122, was introduced into a thermo-sensitive genic male sterile rice line GD-8S, which possessed good grain quality but high susceptibility to rice blast, by using backcross breeding and molecular marker-assisted selection. Five elite improved male sterile lines, RGD8S-1, RGD8S-2, RGD8S-3, RGD8S-4 and RGD8S-5, were selected based on the results of molecular marker analysis, spikelet sterility, recovery rate of genetic background and agronomic traits. Thirty-three representative blast isolates collected from Guangdong Province, China were used to inoculate the improved lines and the original line GD-8S artificially. The resistance frequencies of the improved lines ranged from 76.47% to 100%, much higher than that of the original line GD-8S （9.09%）. On the agronomic characters, there were no significant differences between the improved lines and GD-8S except for flag leaf length and panicle number per plant. The improved lines could be used for breeding hybrid rice with high blast resistance.

Inheritance of the Male Sterility in a New Photo/Thermo-Sensitive Genie Male Sterile Line B06S of Rice

The major male sterile genes in a new photo/thermo-sensitive genie male sterile (PTGMS) line B06S of rice were analyzed by the manipulation of mixture distribution theory. The results indicated that a pair of major male sterile nuclear genes with large effects were responsible for controlling the male sterility of B06S.

Factors Influencing the Outcrossed Seed-Setting Obstacles of Photo-thermo Sensitive Genie Male Sterile Rice

Using photo-thermo sensitive genie male rice (PTGMS) Pei' ai 64S, W7415S, W6154S, N26S, Annong S, Nongken 58S, 7001S and 5088S as female parents and conventional indica lines 8258 and U89 as male parents, the factors affecting outcrossed seed-setting were analyzed. The PTGMS had obstacles in outcrossed seed setting influenced by inheritance and environment at varying degrees. Environmental temperature was regarded as the main factor that resulted in the outcrossed seed-setting obstacles. The sensitive stage was at the early stage of grain filling for outcrossed seed setting. There existed remarkable differences at the sensitivity stage, the duration of sensitive period, the sensitive level and the effective level of outcrossed seed-setting obstacles caused by environmental temperature among different PTGMS lines. Therefore, attention should be paid to outcrossed seed-setting obstacles in selection and utilization of PTGMS lines.

Thermo-sensitive genic male sterile (TGMS) rice line developed by population improvement

In this study population improvement or random polycross plus mass selection was adopted with using N8S, Huaizao4, Xiang2B and Zaoyou1 as parents. To help comprehensively improve the important characters such as combining ability, grain quality, resistance, stigma exsertion rate, etc. The stigma exsertion rate of ZhunS, an elite TGMS line, is 78.6%, 72.4% higher than that of V20A. Hybrids produced by crossing ZhunS with R402, Minhui77 and Minhui63 increased the yield by 3.8%～5.8% and decreased grain chalkiness by 82.1%～84.4% compared with those from crossing V20A with the corresponding R lines.

Elongation of the Uppermost Internode for Changxuan 3S,a Thermo-Sensitive Genic Male Sterile Rice Line

Changxuan 3S, a thermo-sensitive genic male sterile （TGMS） rice line with eui gene, is derived from the TGMS rice line Pei＇ai 64S by irradiation with 350 Gy of ^60Co γ-ray. To elucidate the uppermost internode elongation of the TGMS line with eui gene, Changxuan 3S and its parent Pei＇ai 64S were used to study the effects of temperature on panicle exsertion. At 24℃, the uppermost internode of Changxuan 3S elongated the fastest from the 4^th day before flowering to 0 day （flowering）, being 2.1-fold as that of Pei＇ai 64S, whereas it elongated slowly during the 12^th day to the 4^th day before flowering and the 1^st to the 3^rd day after flowering. The uppermost internode of Changxuan 3S exserted from the flag leaf sheath at 22℃, 24℃ and 26℃, and the length of elongated uppermost internode increased with the decreasing temperatures. At 28℃, though the panicles of Changxuan 3S were still enclosed in the leaf sheath, the degree of panicle enclosure was significantly lower compared with Pei＇ai 64S. Cytological studies on Changxuan 3S showed that the uppermost internode elongation was attributed to the increase of cell number and cell elongation, and the latter was more significant. Moreover, the numbers of outermost and innermost parenchyma cells and the cell length of the uppermost internode reduced with the increasing temperatures.

Introgression of Gene for Non-Pollen Type Thermo-Sensitive Genic Male Sterility to Thai Rice Cultivars

For the two-line hybrid rice system, pol en sterility is regulated by recessive gene that responds to temperature. The recessive gene controlling thermo-sensitive genetic male sterility （TGMS） is expressed when the plants are grown in conditions with higher or lower critical temperatures. To transfer tgms gene（s） control ing TGMS to Thai rice cultivars by backcross breeding method, a male sterile line was used as a donor parent while Thai rice cultivars ChaiNat 1, PathumThani 1, and SuphanBuri 1 were used as recurrent parents. The BC2F2 lines were developed from backcrossing and selfing. Moreover, the simple sequence repeat （SSR） markers were developed for identifying tgms gene and the linked marker was used for assisting selection in backcrossing. The identification lines were confirmed by pol en observation. The results showed the success of introgression of the tgms gene into Thai rice cultivars. These lines will be tested for combining ability and used as female parent in hybrid rice production in Thailand.

小麦光温敏核雄性不育系BS366和BS1086杂种一代结实率研究

为了解小麦光温敏核雄性不育系BS366和BS1086的育性恢复状况,评估恢复系人工定向改良对其恢复能力的效果,在不育生态区对以55份定向改良的恢复系和33份非改良恢复系(常规小麦)为父本配制的杂种一代的结实率进行了分析。结果表明,BS366和BS1086杂种一代结实率变化范围为15.42%~140.34%和20.28%~119.07%(国际法),平均结实率分别为73.02%和72.21%,二者差异不显著,说明2份母本的育性恢复能力差异不显著。BS366和BS1086杂种一代结实率主要分布于40%~110%和60%~100%间,BS1086杂种一代结实率分布更为集中。BS366和BS1086与改良父本的杂种一代平均结实率分别为84.45%和78.97%,与非改良父本的杂种一代平均结实率分别为53.97%和60.95%,改良父本的平均恢复力高于非改良父本,表明定向改良有利于提高恢复系的恢复力。BS366和BS1086与相同父本的杂种一代结实率差异在-41.66%~61.93%之间,表明相同父本对不同母本的育性恢复力存在差异。BS366和BS1086与相同改良父本的杂种一代中有61.82%的结实率差异分布于-10%~20%之间,与相同非改良父本的杂种一代中有51.52%的结实率差异在-30%~0%之间,表明同一改良父本对不同母本的育性恢复力差异小于非改良父本。BS366与14份父本、BS1086与7份父本的杂种一代结实率高于对照,其中父本14YH261、14YH551、SD036与2份母本杂种一代的结实率均高于对照,说明具有可用恢复力的父本不仅数量极少,而且对不同母本的恢复力存在较大差异。杂种一代主茎穗的平均结实率极显著大于分蘖穗。因此,建议对不同母本分别开展强恢复力的恢复系筛选,并以中选强恢复系为亲本进行定向改良,有效增加杂种一代结实率达对照水平的组合数量,为获得更多强优势杂交组合奠定基础。

外源茉莉酸甲酯对BNS366雄性育性的影响

为探究外源茉莉酸甲酯(methyl jasmonate,MeJA)对小麦温敏雄性不育系BNS366雄性育性的影响,以BNS366及其近等基因系郑麦366为试验材料,在正常秋季播种(2020年10月12日)和晚播(2020年12月2日)条件下,分别于2021年3月22日和3月31日开始至开花期前,设置0(清水,对照)、50、100、200、300、400和500μmol·L^(-1) MeJA(水溶液,喷施)7个处理,比较分析了不同处理间花粉可育率和自交结实率的差异。结果表明:在正常秋季播种条件下,郑麦366花粉可育率、国内法自交结实率和国际法自交结实率分别为86.31%、70.36%和112.22%,在晚播条件下分别为82.53%、92.53%和166.18%,均正常可育;在正常秋季播种条件下,200μmol·L^(-1) MeJA处理的国际法自交结实率为70.15%,比对照显著降低了42.07个百分点;在晚播条件下,200、300和500μmol·L^(-1) MeJA处理的国内法自交结实率分别为74.71%、74.01%和73.45%,比对照显著降低了17.82、18.52和19.08个百分点;在两个播期下,郑麦366其他浓度MeJA处理的上述3个指标与对照差异均不显著。在正常秋季播种条件下,BNS366花粉可育率为零,达到全不育水平,国内法自交结实率和国际法自交结实率均为0.24%,100μmol·L^(-1) MeJA处理的花粉可育率为88.25%,达到正常可育水平,国内法自交结实率和国际法自交结实率分别为56.41%和94.01%,能正常结实,与对照差异达到显著水平;在晚播条件下,BNS366的上述3个指标分别为51.72%、41.23%和93.08%,正常可育,100μmol·L^(-1) MeJA处理的国际法自交结实率为39.72%,比对照显著降低了53.36个百分点;在两个播期下,BNS366其他浓度MeJA处理的上述3个指标与对照差异均不显著。由此说明,在2020-2021年,外源MeJA对郑麦366和BNS366可育植株的雄性育性可能具有降低效应,对BNS366不育植株的雄性不育性具有较强恢复效应。

光温敏两系不育系‘稆S’的选育与应用

[目的]选育优良的光温敏两系不育系,以提高其杂种后代的产量和品质。[方法]采用光温敏两系不育系‘RGD-7S’为母本与三系保持系‘中浙B’杂交,经5年9代的系谱法选育,育成不育性稳定、稻米品质优、异交特性好、株叶形态优良的光温敏两系不育系‘稆S’,并分析了其主要特征特性及杂种优势。[结果]‘稆S’不育性稳定,育性转换临界温度<23℃,稳定不育期127 d左右。该不育系制种产量高,异交特性好,柱头外露率78.62%,其中双边外露率39.74%。采用‘稆S’配组的高产、优质两系杂交稻组合‘稆两优质美’先后通过了福建省和海南省农作物品种审定。[结论]光温敏两系不育系‘稆S’不育性稳定,育性转换临界温度低,稻米品质优,配合力强,应用前景广泛。

MAPPING OF THE RICE (ORYZA SATIVA L.) THERMO-SENSITIVE GENIC MALE STERILE GENE TMS5 WITH EST AND SSR MARKERS

With the cDNA suppression subtraction hybridization method, a spikelet-specific cDNA library was constructed that expressed at meiosis stage in rice. A total of 121 cDNA fragments were selected from the library and used as EST (expressed se-quence tags) markers to detect the polymorphism between Annong N, a normal fertile Indica rice line and Annong S-1, its spontaneous mutant with thermo-sensitive genic male sterility, using the RFLP (restriction fragment length polymorphism) technique. HN57, one of the EST probes, could detect poly-morphism between them. The results of segregation analysis with the F2 population developed from An-nong S-1 and Annong N indicate that HN57 co-seg- regates with the thermo-sensitive genic male-sterility controlled by tms5, the recessive gene in Annong S-1. This marker is located on the 31.2-cM region of the chromosome 2 of RGP (rice genome research pro-gram) genetic map. To further determine the location of tms5, 80 SSR (simple sequence repeat) markers around this region were developed, and 12 of them were polymorphic. And finally, the tms5 was mapped within region of 181 kb by using these new markers.

Detection and Analysis of High Temperature Sensitivity of TGMS Lines in Rice Using AMMI Model

With the AMMI (additive main effects and multiplicative interaction) analysis model, thedetermination of the sensitivity to temperature among different TGMS (thermo-sensitivegenic male sterile) lines was performed. To assess the genetic differences due to hightemperature stress at the fertility-sensitive stage (10-20d before heading), sevengenotypes (six TGMS lines and the control Pei-Ai64S) were grown from May 4 at sevendifferent stages with 10d intervals. The temperatures at the fertility-sensitive stagesinvolved twelve levels from<20 to>℃ under the regime natural conditions in Hangzhou,China. There was considerable variation in pollen fertility among genotypes in responseto high temperature. Five genotypes identified as TGMS lines as their percentages offertile pollens were lower than or close to that of the control except for the unstableline RTS19 (V6). When the temperatures at the fertility-sensitive stage were at Ⅰ-Ⅳ,Ⅴ-Ⅵ and Ⅶ-Ⅻ, the percentages of fertile pollens varied in the ranges of 46.46-48.49%,19.62-22.79% and 3.49-5.87%, respectively. The critical temperatures of sterility andfertility in the five TGMS lines were 25.1 and 23.0℃, respectively. Considering theamounts and directions of main effect and their IPCA (interaction principal componentsanalysis), we can classify the lines and temperature levels into different groups, anddescribe the characteristics of genotypetemperature interaction, offering the informationand tools for the development and utility of thermo-sensitive male sterile lines.Several TGMS rice lines with their reproductive sensitivity to high temperature that canbe screened using the AMMI model may add valuable germplasm to the breeding program ofhybrid rice.

Fertility Expression of TGMS-Genes in the Backgrounds of indica CMS-lines,B-lines and R-lines of Hybrid Rice

The generation fertility of 51 F1, 19 F2 and 6 BC1 between 3 thermo-sensitive genic male sterile lines （TGMS-lines） Pei＇ai 64S, 6311S and 360S, and the three lines of hybrid rice including 7 indica cytoplasmic male sterile lines （CMS-lines） and their corresponding maintainer lines （B-lines） and 3 indica restorer lines （R-lines） were investigated to study the expression of TGMS-genes in the backgrounds of the three lines of hybrid rice. Pei＇ai 64S has stronger fertility restoring （Rf） genes for CMS-lines and its TGMS trait is governed by 2 pairs of independent recessive genes; The TGMS trait of 6311S is governed by a single recessive gene with weaker Rf-gene in 6311S and the TGMS trait of 360S is governed by a single recessive gene with no Rf-gene in 360S. The investigation on the fertility of F1 plants between 5 CMS-lines and 4 TGMS generations selected from F2 plants of 4 CMS-lines x 6311S confirmed that the expression of TGMS-gene was controlled by Rf-gene in the genetic background of cytoplasm of CMS-lines, but not affected by Rf-gene in the genetic background of normal fertile cytoplasm. The potential breeding strategies of TGMS-lines with cytoplasm of CMS-lines and CMS-lines with the nucleus of TGMS-genes were discussed.

不同水稻tms5突变位点对雄性不育起点温度的影响

【目的】研究不同水稻tms5突变位点对雄性不育起点温度的影响,探讨不育起点温度遗传调控途径。【方法】在水稻TMS5的6个外显子上设计11个CRISPR/Cas9基因编辑靶点,依次命名为T501—T511,构建相应载体转化粳稻品种日本晴和籼稻品种明恢86,获得各靶点的tms5移码突变体。田间自然高温及人工气候箱(设日平均22、24和28℃3种温度)条件下分析tms5突变体的花粉碘染及自交结实率,鉴定不育起点温度。【结果】粳稻日本晴tms5突变体的不育起点温度高于28℃,籼稻明恢86的不同tms5突变体不育起点温度为22~28℃。此外,同一遗传背景下,通过T501靶点编辑产生的tms5-1移码突变体不育起点温度均显著高于T502靶点的tm5-2突变体,其他位点上的tms5突变体育性特征与tm5-2突变体并无差异。基因表达量分析表明,tms5-1突变体幼穗Ub_(L40)基因表达量显著低于tm5-2突变体的表达量。【结论】水稻tms5突变体不育起点温度不仅受遗传背景的影响,tms5基因突变位点不同也会影响不育起点温度,特别是T501位点与其余位点突变体间不育起点温度差异显著,为研究水稻tms5两系不育起点温度的分子机理及遗传调控网络提供了新思路。

广适籼型两系不育系7011S的选育与应用

7011S系安徽省农业科学院水稻研究所以生产上广泛使用的籼型两系不育系1892S为母本,与抗稻瘟病材料谷梅4号杂交、自交,经5年7代选育而成的籼型温敏核不育系。该不育系具有株型适中、叶色深绿、分蘖力较强、茎秆较粗、不育起点温度低、异交结实率高、配合力强及广适等特性,2019年8月通过安徽省农作物品种审定委员会鉴定。以7011S为母本、R202为父本配制的两系杂交籼稻新组合两优7002具有优质、高产、抗逆特点,于2022年通过国家农作物品种审定委员会审定。

水稻籼型光温敏核不育系福7112S的选育及应用

籼型光温敏核不育系福7112S是以1892S为母本与优质自育材料R0564杂交,采用系谱法选育而成的。该不育系育性稳定、败育彻底、株型紧凑、抗性较好、米质较优,2019年通过安徽省农作物品种审定委员会组织的技术鉴定。本文介绍了福7112S的选育过程,阐述了其主要的特征特性和育性表现,分析了其配组应用,总结了繁殖制种技术,以期为福7112S的推广与应用提供参考。

优质籼型水稻光温敏核不育系‘7-163S’特征特性及其基因型解析

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