Genetic Analysis and Primary Mapping of pms4, a Photoperiod-Sensitive Genic Male Sterility Gene in Rice (Oryza sativa)

To understand the genetic characteristics of a new photoperiod-sensitive genic male sterile line Mian 9S, some reciprocal crosses were made between Mian 9S and six indica rice materials, Yangdao 6, Luhui 602, Shuihui 527, Mianhui 725, Fuhui 838 and Yixiang 1B. Genetic analysis results suggested that the photoperiod-sensitive genic male sterility （PGMS） of Mian 9S was controlled by a single recessive nuclear gene. Thus, the F2 population derived from the cross of Yangdao 6/Mian 9S was used to map the PGMS gene in Mian 9S. By using SSR markers, the PGMS gene of Mian 9S was mapped on one side of the markers, RM6659 and RM1305, on rice chromosome 4, with the genetic distances of 3.0 cM and 3.5 cM, respectively. The gene was a novel PGMS gene and designated tentatively as pms4. In addition, the application of the pms4 gene was discussed.

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.

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.

Preliminary Study on Inheritance of Stigma Exertion in Wheat Thermo-photo Sensitive Genic Male Sterile Line

Stigma exertion is one of the key factors for improving the outcrossing ability of wheat thermo-photo sensitive genic male sterile（TPSGMS） line. A DH population derived from K239S/K92 S was constructed to investigate the inheritance of stigma exertion. K239 S and K92 S are TPSGMS lines with higher and lower stigma exertion rates（SER）, respectively. The SERs of parents, reciprocal crosses and the DH population were evaluated for two consecutive years. The results showed that no significant difference was observed in SER between F1 s of K239S/K92 S and K92S/K239 S,implying that stigma exertion was a trait controlled by nuclear gene（s）. In the DH population, the segregation of low and high SERs fitted to a ratio of 3 ∶1 by Chisquare test, suggesting that the stigma exertion of K239 S was controlled by one pair of recessive genes. In addition, the effects of temperature and humidity on the expression of stigma exertion were also discussed.

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.

Isolation and Analysis of the Promoter of OsRacD from PhotoperiodSensitive Genic Male Sterile Rice

By using OsRacD cDNA as probe to screen the genomic library of photoperiod sensitive genic male sterile rice line Nongken 58S, a positive clone containing 2 kb promoter and 396 bp coding region of OsRacD was obtained. Compared with the promoter of OsRacD cloned by reverse PCR from normal rice variety Nongken 58 （Nongken 58N）, the homology was 99.8%, and the different nucleotides were outside the predicted response elements in promoter, suggesting that the fertility between rice varieties Nongken 58S and Nongken 58N under the long-day conditions was not attributed to the difference in the structure of OsRacD upstream regulation sequences, but to the developmental regulation of gene differential expression.

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.

Plant Temperature for Sterile Alteration of a Temperature-Sensitive Genic Male Sterile Rice,Peiai64S

The forecast of sterile alteration for the temperature-sensitive genic male sterile （TGMS） line in two-line hybrid rice seed production was traditionally based on screen temperature determined by weather station. The article put forward a new approach based on plant temperature, which was more exact and direct than the traditional method. The result of the simulation of the self-seeded setting rate of a widely used TGMS line, Peiai64S, by several temperature parameters and durations, showed that the fertility was directly affected by the plant temperature at a height of 20 cm or the air temperature around it in three days duration. Using the stem temperature of three days at a height of 20 cm as the simulation parameter, the fertility of Peiai64S had the maximum, minimum and optimum temperatures as 22.8, 21.7 and 22.5℃, respectively, whereas 23.2, 21.5 and 21.8℃ when using the air temperature of three days around the height of 20 cm as the parameter. Such temperature indices can be used to conclude the sterile alteration of TGMS for safeguarding seed production of twoline hybrid rice. The article also established a statistic model to conclude plant temperature by water temperatures at inflow and outflow, and air temperature and cloudage from weather station.

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.

Progresses and Strategies of Gene Mapping and Isolating of Photo(thermo) sensitive Genic Male Sterile Rice

Considering the research on classical genetics of photoperiod(therm) sensitive genic male sterile rice, it is important to select the sterile lines and their segregating population controlled by one pair of gene in mapping and isolating sterile genes. It is discussed the advantages, disadvantages and the reasons leading to various mapping results of chromosome location of sterile genes through gene marker, isozyme marker and DNA marker techniques. In comparison to isolation of photo(thermo) sensitive sterile genes via various plant gene cloning techniques, it was concluded that map based cloning was acceptable, but it is still difficult to locate the loci of sterile genes within 1cM. On the other hand, “sensitivity to environment”, an important characteristic of sterile lines can be fully utilized by DD PCR and (or) RDA techniques. Therefore, these two techniques were considered as the effective ways to isolate sterile genes.

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.

Identifying and mapping cDNA fragments related to rice photoperiod sensitive genic male sterility

The differentially expressed cDNA fragments have been obtained by differential screening with cDNA-RAPD technique in photoperiod sensitive genic male sterile (PGMS) rice. Some of them have been reassessed with Northern blot hybridization, from which a PGMS-related positive fragment, RPG43, has been identified. Further analysis on RPG43 with Southern blot and RAPD indicates that the fragment is a single-copy sequence and its mRNA has been processed after transcription. Sequence analysis reveals that RPG43 is 744 bp in length and contains a 60 bp region (from 126th to 185th bp) showing 72% homology to a human DNA sequence, pac pDJ-356d6, on chromosome 11. So it is a new sequence found in plant and its GenBank access number is AF126027. In addition, RPG43 has been mapped to a position 3.8 cM away from RFLP marker R1553 on chromosome 5 of rice.

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。

香型温敏核不育系皖香22S的选育研究

皖香22S是安徽省农业科学院水稻研究所以1892S为母本、湖南湘穗种业有限责任公司引进的香型籼稻亲本常香39S为父本进行杂交选育的香型温敏核不育系,2023年8月通过安徽省技术鉴定。皖香22S株高85.0 cm,平均每穗总颖花数193.5粒,千粒重24.0 g,株型紧凑,分蘖力较强,叶片直立内卷,有香味,冷水池育性鉴定在14.5 h、23.5℃、6 d条件下,花粉不育度99.95%,套袋自交结实率0.25%。皖香22S在合肥5月20日播种,播始历期为84 d,比近似品种常香39S短5 d左右,主茎叶龄13.2叶。柱头外露率和异交结实率较高。稻瘟病、稻曲病和白叶枯病抗性较好;米质较优;所配组合杂种优势强,其中皖香22S/银月丝苗在2023年的小区优势鉴定中,较丰两优4号增产9.35%,差异达极显著水平。

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。

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.

光/温敏核不育系的不育机理及两系杂交稻的发展与展望

本文回顾了中国两系杂交水稻的发展历史;介绍了两系法中光/温敏型核不育系遗传资源的发现和选育;总结了植物中光温敏型雄性不育的遗传学和细胞学机制;并对水稻两系法中光/温敏型雄性不育系的遗传资源优化及育种策略提出了建议。

优质籼型光温敏核不育系荃时S的选育与应用

介绍了荃时S的选育过程、特征特性、配组应用及繁殖技术要点。荃时S是以银312S为母本,与配合力好的优良恢复系YR0822杂交,采用系谱法选育而成的籼型光温敏核不育系,具有育性转换起点温度较低、不育性稳定、异交率高、抗倒伏、米质优等特点,2021年通过安徽省农作物品种审定委员会组织的专家技术鉴定。该不育系与恢复系银泰香占配制的籼型两系杂交水稻品种时两优泰香具有中抗稻瘟病、米质达部标优1级、丰产等特点,并获得2022年(第四届)安徽省优质稻品种食味品质籼稻金奖,2022年通过国家农作物品种审定委员会审定。

籼型特色水稻两系不育系红17S的选育

红17S是福建省南平市农业科学研究所以广占63S为母本与南平当地优质特色红米美人红杂交,后又与籼型两系不育系K12S复交,经福建建阳及海南三亚等地充分利用自然生态条件,4 a 8代培育成的特色红米温敏核不育系,2022年通过福建省农作物品种审定。总结了红17S的选育过程、配组应用及配套繁殖、制种技术。