Characterization of Reverse Thermo-sensitive Genic Male Sterile Lines in Cold Regions of China

Yannong S, a reverse thermo-sensitive genic male sterile （TGMS） line, is sterile at high temperature and fertile at low temperature. In the present study, the fertility of Yannong S and the japonica reverse TGMS lines derived from it was in- vestigated in Harbin, Heilongjiang from 2013 to 2015. The results showed that Yan- nong S was completely male sterile in July and August （the hottest months of the year in Harbin）, and the seed setting rate of bagged panicles was 0. We also in- ferred that the critical temperature for inducing the sterility of Yannong S was higher than 28 ℃. Most of the japonica reverse TGMS lines derived from Yannong S were completely aborted in July and August, with a seed setting rate of 0. It can be con- cluded that the reverse TGMS lines derived from Yannong S are genetically stable lines that have a long period of sterility in cold regions, so they may be the ideal genetic materials for the development and in-situ seed production of japonica hybrid rice.

Characterization and fine mapping of RTMS10, a semi-dominant reverse thermo-sensitive genic male sterile locus in rice

The discovery and application of environment-sensitive genic male sterile(EGMS) rice germplasm provide an easy method for hybrid rice breeding and have made great contributions to hybrid rice production. Typically, the photoperiod-and thermosensitive GMS(P/TGMS) lines utilized in two-line hybrid systems are male sterile under long day or/and high temperature but fertile under short day or/and low temperature conditions. However, Yannong S(Yn S), a reverse TGMS(rTGMS) line, is sterile under low temperature(<29℃) and fertile under high temperature(>29.5℃). Here, we report a genetic study on the rTGMS trait in Yn S. Interestingly, the F1 plants of the cross between Yn S and a cultivar, L422, were male sterile at 22℃ and completely fertile at 27℃. Moreover, the segregation ratio of fertile and sterile individuals in Yn S/L422 F2 populations changed from 1:3.05 to 2.95:1 when the ambient temperature increased, showing that the rTGMS trait exhibits semidominance in Yn S. We further found a locus on chromosome 10, termed RTMS10, which controls the rTGMS trait in Yn S. We then finely mapped RTMS10 to a ~68 kb interval between markers ID13116 and ID1318 by Yn S/L422 BC6 F2 populations. A near iso-genic line(NIL) NL1 from the BC6 F3 generation was developed and the pollen of NL1 became abnormal from the meiosis stage under low temperature. In summary, we identified an rTGMS locus, RTMS10, and provided co-segregated markers, which could help to accelerate molecular breeding of rTGMS lines and better understand the rTGMS trait in rice.

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.

Identification and fine mapping of rtms1-D,a gene responsible for reverse thermosensitive genic male sterility from Diannong S-1X

Thermosensitive genic male sterility(TGMS)has been widely used in two-line hybrid rice breeding.Due to hybrid seed production being highly affected by changeable environments,its application scope is limited to some extent.Thus,it is of great importance to identify potential TGMS genes in specific rice varieties.Here,Diannong S-1 xuan(DNS-1 X),a reverse TGMS(RTGMS)japonica male sterile line,was identified from Diannong S-1.Genetic analysis showed that male sterility was tightly controlled by a single recessive gene,which was supported by the phenotype of the F;and F;populations derived from the cross between DNS-1 X and Yunjing 26(YJ26).Combining simple sequence repeat(SSR)markers and bulked segregation analysis(BSA),we identified a 215 kb region on chromosome 10 as a candidate reverse TGMS region,which was designated as rtmsl-D.It was narrower than the previously reported RTGMS genes rtmsl and tms6(t).The fertility conversion detected in the natural environment showed that DNS-1 X was sterile below 28-30℃;otherwise,it was fertile.Histological analysis further indicated that the pollen abortion was occurred in the young microspore stage.This study will provide new resources for two-line hybrid rice and pave the way for molecular breeding of RTGMS lines.

Selection of Submergence Tolerant Homozygous Line by STS Marker and Twice Submergence Stress

One sequence tagged site marker Subl-1 and twice submergence stress method were used in selection of submergence tolerant homozygous line from Sub-lBS, a submergence tolerant, bentazon sensitive and photoperiod-sensitive and/or thermo-sensitive genic male sterile line that developed by our laboratory. The results revealed that the original Sub-lBS was heterozygous in SublA-1 locus even though it was identical in almost all of agronomical traits and the segregation of SublA-1 was in accordance with Mendelian law based on chi-square test. And then the original Sub-IBS was divided into two groups： one was ofSublA-1 introgression and the other was not; and the two groups were tested by twice submergence stress method. After the first submergence stress that lasted for 12 d, the average plant heights were significant difference at the 1% level between the two groups. After recovery for 10 d, the second submergence stress sustained for 18 d was carried on; and the group with SublA-1 gene was found apparently tolerant than the other group in submergence tolerance.

聚合水稻温敏核不育基因和反温敏核不育基因创制永久核不育系

【目的】制种安全是影响两系杂交稻持续健康发展的关键问题。本研究旨在探索解决两系杂交水稻的制种安全隐患途径。【方法】以16个(光)温敏核不育系与4个反温敏核不育系配组并对F1育性进行观察,选择矮紫S(温敏核不育系)和矮雁s(反温敏核不育系)为构建永久核不育系的供源亲本。通过回交将矮紫S和矮雁s的育性基因导入到桥梁亲本天丰B中,分别育成天丰B的近等基因系天丰S和天丰s。【结果】天丰S和天丰s杂交获得的天丰Ss即为永久核不育系。对天丰Ss的育性、可恢性等特性的研究表明,天丰Ss在自然长日高温和短日低温下均表现不育;天丰Ss及其3个亲本与5个恢复系杂交的F1组合结实率无明显差异。【结论】永久核不育系的创制有望从根本上解决两系杂交水稻的制种安全问题。

Studies on GA_3 Spraying Dosage for eui TGMS Rice Changxuan 3S in Its Hybrid Seed Production

[Objective] Changxuan 3S was thermo-sensitive genicmale sterile（TGMS）rice selected from irradiated seeds of Peiai 64S by 350 Gy^（60）Coγ-ray.The aim of the study was to confirm GA3 spraying dosage of Changxuan 3S with eui gene in its hybrid seed production.[Method] Changxuan 3S possessing eui gene and its parent Peiai 64S were chosen as materials.Comparison studies on sensitivity to GA3 in their hybrid seed production were carried out.[Result] The suitable stage for spraying GA3 in the hybrid seed production of Changxuan 3S was at 10% of panicles headed;The optimal dosage was 90 g/hm2 with 2 split sprayings,the first spraying of 45 g/hm2 at heading of 10% panicles and the second one of 45 g/hm2 on the following day.Under the condition of spraying GA3 at the rate of 90 g/hm2,the panicle neck exsertions of Changxuan 3S was ＋1.78 cm,and exserted stigma rate and seed setting rate of Changxuan 3S were 96.87% and 36.44%,being 21.46% and 16.33% more than those of Peiai 64S,respectively.The theoretical yield of ＂Changxuan 3S/9311＂ reached 2 931.90 kg/hm2,which was increased by 1 259.40 kg/hm2 comparing with ＂Peiai 64S/9311＂.[Conclusion] Compared with Peiai 64S,Changxuan 3S is more sensitive to GA3,which results in no or little using GA3 in seed production of Changxuan 3S.Moreover,Changxuan 3S showed higher yield potential than Peiai 64S.

野栽远缘杂交来源的反向温敏核雄性不育系不育性遗传研究

利用3个来源于野生稻与栽培稻杂交后代的反向温敏不育系R6S、N13S、Tb7S为材料,开展了反向温敏不育系不育性遗传研究,结果表明反向温敏不育系N13S的育性是细胞核内1对隐性基因控制的;Tb7S的育性是受隐性核基因控制的,F2代的育性分离比为9∶7,不育性状表现为2对基因的独立遗传;R6S的F2代育性分离比为37∶27,受细胞核内的3个隐性基因位点分别位于不同染色体上而通过互补作用的独立遗传.为进一步分离、定位及克隆有关这些反向温敏不育基因及发展分子标记辅助选择选育反向温敏不育系奠定了良好的遗传学基础.

大豆反向光敏感细胞核雄性不育突变体88-428BY育性特征分析

为明确大豆光敏感细胞核雄性不育突变体88-428BY不育的生理生化因素,本研究对其总花芽进行能量物质和抗氧化酶活性检测,结果表明88-428BY不育株的可溶性糖、淀粉和ATP含量均显著低于可育株,说明不育株中为花药生长发育提供能量的物质在代谢过程受到抑制最终影响其育性。抗氧化物酶超氧化物酶(SOD)、过氧化氢酶(CAT)和抗坏血酸过氧化物酶(APX)活性在不育株中均低于可育株,说明88-428BY败育的原因与抗氧化物酶活性也有一定关系。总结以上结果推测,大豆光敏感细胞核雄性不育突变体88-428BY不育可能与花药发育过程相关的能量物质或者酶活性等有关,为后期深入研究大豆光敏不育机理和推进两系杂交大豆育种提供理论佐证。