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.

Mapping of a gene for photoperiod-sensitive genic male sterility in Nongken 58s on chromosome 5 of rice

A photoperiod-sensitive genic male sterile (PGMS) rice was found in 1973 as a spontaneous mutant of Nongken 58, a japonica variety. Pollen fertility of Nongken 58s (N58s) is completely sterile when grown under long-day conditions, whereas fertile under short-day conditions. This PGMS was found to be controlled by one or two recessive gene(s), of which one gene(pms)was linked to a marker gene(d-1) on chromosome 5. In order to identify a more precise location of the pms, we analyzed the populations of BCFand BCFof N58s//N58s/KL211(v-10, virescent) and N58s//N58s/KL520 (gh-1, gold hull). The marker genes v-10 and gh-1 are located on the flanking region of d-1. The F, plants of two crosses were fertile. The number of fertile and sterile individuals in BCFfit

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.

Differential Expression Analysis of Genic Male Sterility A／B Lines in Chinese Cabbage—Pak—Choi（Brassica campestris ssp.chinensis Makino）

To determine differential expression of genie male sterility A/B lines in Chinese cabbage-pak-choi (Brassica campestris ssp. chinensis Makino var. communis Tsen et Lee), we used the RNA fingerprinting technique, cDNA-AFLP analysis, in different developmental stages and different tissues. While no obvious differential expressions were observed in rosette leaves, florescence leaves, and scapes, some differential expressions were found in alabstrums of A/B lines and among leaves, scapes and alabstrums. We analyzed the al-abstrums collected in different developmental stages with 10 primer combinations. We got a unique band between middle size alabstrums and large alabstrums in B line in one of the ten pair primers, and in another one pair, one band reflecting a higher gene-expression level in A line than that in B line was obtained. No unique bands were found with the other primer combinations. The bands reflecting different gene-expression level were confirmed by Northern hybridization. The results indicated that cDNA-AFLP was a suitable tool for studying differential expression of genie male sterility in plants. SDS-polyacrylamide gel electrophoresis patterns of soluble proteins further verified the difference in A/B lines.

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.

Studies on the Photoperiod Sensitive Characters of Male Fertility Alteration of Peiai64S' Main Male Genic Sterile Gene

Peiai64S, an indica male sterile rice with a male fertility alteration under different environments, is selected from the offspring of indica rice crossed with Nongken58S. Nongken58S, a japonica pho-toperiod sensitive genie male sterile rice (PGMS), deriving from a natural mutant plant individual of normal japonica rice variety, Nongken58, is used as a male sterile gene donor of Peiai64S. But Peiai64S is not a typical PGMS rice, the male fertility is sensitive to temperature just as thermo-sensitive genie male sterile rice (TGMS). We have selected typical PGMS plants in F2 population of Peiai64S× Nongken58, whose ratio of fertile plants to sterile plants is nearly 3:1. The sterility inheritance conformed to one pair of gene segregation model. The result indicates the main male sterile gene in Peiai64S is not other than the PGMS gene, and comes from Nongken58S. The genetic background affects effective expression of the PGMS gene. This suggests that we ought to focus on optimizing the genetic background of the PGMS gene in PGMS rice breeding, and select an ideal genetic background as a transgenic background in molecular breeding.

pms3 is the locus causing the original photoperiod-sensitive male sterility mutation of‘Nongken 58S'

Photoperiod-sensitive genic male sterile (PSGMS) rice is a very useful germplasm for hybrid rice development. It was first found as a spontaneous mutant in a japonica cultivar ’Nongken 58’. pms3 on chromosome 12 was determined to be the locus where the original PSGMS mutation occurred, changing the normal cultivar Nongken 58 to PSGMS Nongken 58S. Large amounts of RAPD and AFLP analyses were also conducted for the fine mapping of the pms3 genomic region, which resulted in 4 molecular markers linked to pms3. Although these markers somewhat increased the marker density of this region, the pms3 locus is still located in a marker-sparse region.

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.

Selection and Characterization of a Novel Photoperiod-Sensitive Male Sterile Line in Upland Cotton

Upland cotton （Gossypium hirsutum L.） shows strong heterosis. However, heterosis is not widely utilized owing to the high cost of hybrid seed production. Creation of a photoperiod-sensitive genetic male sterile line could substantially reduce the cost of hybrid seed production in upland cotton. Such a mutant with virescent marker was found by space mutation in near-earth orbit and its traits had been stable after 4 years of selection in Anyang and Sanya, China. This mutant was fertile with an 11-12.5 h photoperiod when the temperature was higher than 21.5 ℃ and was sterile with a 13-14.5 h photoperiod. Genetic analysis indicated that both traits were controlled by a single recessive gene or two closely linked genes. Also, the cytological observations and transcriptome profiling analysis showed that the degradation of pollen grain cytoplasm should be the primary reason why the mutant line were male sterile under long-day conditions.

Identification of differentially expressed genes in photo-period sensitive genic male sterile rice by randomly amplified cDNAs using RAPD primers

The fertile and sterile young panicle representational populations were constructed by bulked sampling method using young panicles in the photo-period sensitive stage of fertility transformation.Two populations were analyzed using cDNA-RAPD. The results showed that: ( ⅰ ) bulked sampling method can be employed to analyze differentially expressed genes using cDNA-RAPD, taking an advantage in avoiding false positive caused by conventional sampling method. (ⅱ) Among 150 random primers used, 83 primers amplified the same banding patterns, and 34 primers amplified the same banding pattern but different staining of intensity on gel. ( ⅲ ) 33 primers amplified differential cDNA bands between fertile and sterile cDNA populations, and the ratio of polymorphism was 22%. It is concluded that there may exist a lot of genes relating to sterility, which makes the differentially expressed cDNA fragments complicated.

Construction and characterization of a bacterial artificial chromosome library of thermo-sensitive genic male-sterile rice 5460S

In order to develop a detailed physical map of the thermo-sensitive genie male-sterile (TGMS) gene-encompassing region and finally clone the TGMS gene, a high-quality rice bacterial artificial chromosome (BAC) library from TGMS rice 5460S was constructed. The method of constructing BAC library was examined and optimized. The 5460S library consists of 19 584 BAC clones with an average insert size of 110 kb, which represents about 5 times rice haploid genome equivalents. Rice inserts of up to 140 kb and 250 kb were isolated and appeared stable after 100 generations of serial growth. Hybridization of BAC clones with mitochondrial and chloroplastic genes as probes demonstrated that this library has no organellar contamination. The 5460S library was screened with 3 molecular markers linked to tmsl gene as probes and at least 1 BAC clone was identified with each probe. The insert ends of positive clones were successfully isolated using thermal asymmetric interlaced PCR (TAIL-PCR) technique.

Molecular tagging of a genic male-sterile gene in rice

The sterility of Pingxiang male-sterile rice (Pins), possibly derided from a spontaneous mutation in Pingxiang fertile rice (Pmf), was previously reported to be controlled by a single dominant nuclear gene. It can be restored to fertility either by a dominant epistatic gene or by higher temperature treatment at the early stage of inflorescence development. In order to tag the genie male-sterile gene, Pms, Pmf and Ce 64, a cytoplasmic male-sterile restoring line without the epistatic gene for Pms, were used to construct mapping populations. Two segregation populations, "(Pms/Ce 64) F1s (sterile plant)//Pmf" F1 and "Pms//(Pmf/Ce 64) F1" F1, were simultaneously developed. Subsequently, the genie male- sterile gene was mapped between a simple sequence length polymorphism marker, RM228, and a restriction fragment length polymorphism marker, G2155, with distances of 14.9 and 2.6 cM, respectively. The tagged dominant genie male-sterile gene is temporarily designated Ms-p.

甘蓝型油菜细胞核+细胞质雄性不育三系的研究与利用 Ⅳ.隐性细胞核+波里马细胞质雄性不育三系的创建

甘蓝型油菜隐性细胞核雄性不育与波里马细胞质雄性不育可能具有完全不同的不育基因系统;采用有性杂交和连续回交的方法,将甘蓝型油菜隐性细胞核不育基因导入到含有波里马不育细胞质的基因型中,并得到甘蓝型油莱隐性细胞核+波里马细胞质雄性不育系RGCMS—S45A和RGCMS—117A及其相应的保持系S45B和117B;采用测交方法,从隐性细胞核雄性不育系中筛选出甘蓝型油菜隐性细胞核+波里马细胞质雄性不育保持系9个;波里马细胞质雄性不育的恢复系均是隐性细胞核+波里马细胞质雄性不育的恢复系。

三个甘蓝型油菜隐性细胞核雄性不育系小孢子发生的细胞学研究

对三个甘蓝型油菜隐性细胞核雄性不育系花药发育的解剖学观察表明，S45A和117A小孢子发生的细胞学特点相同，其雄性败育于减数分裂后的四分体至单核小孢子阶段。其主要特点是减数分裂完成后，小孢子不形成花粉壁并开始解体；同时，绒毡层细胞径向扩大和小液泡化并逐渐解体。9012A雄性败育于花粉母细胞至减数分裂四分体阶段。其主要特点是花粉母细胞减数分裂后胼胝质壁不降解和不释放出四分体，并且四分体开始液泡化解体；同时，绒毡层细胞逐步大液泡化，直至最后解体。

水稻光敏核不育系N422s的广亲和性及分类地位初探

N422s是以晚粳型光敏核不育系7001s为供体,广亲和系轮回422为受体,经过杂交转育而成的稳定的中粳型光敏核不育系。亲和性研究表明,N422s不仅与4个标准测验种杂交产生具有正常育性的F_1代,而且与30个不同类型的地方品种(系)杂交F_1代也表现较高的育性。因此,该不育系具有广亲和性及配组自由度大的特点。N422s在形态指数及酚反应上表现为粳型,在杂交亲和力上也偏粳。但是,选用与籼粳分类有关的多位点同工酶进行分析,结果表明它属于籼稻。在光温生态反应上,它也表现出与原始亲本培迪(Padi)相似的反应型。考虑到N422s的广亲和性来自印度尼西亚的平原地方品种培迪,在分类地位上它归属于粳亚种下的爪哇群。

应用花培技术选育优质低温敏水稻核不育系金山S-1

选用性状互补的两个亲本8-1S和香125S杂交,采用系谱育种和花药培养相结合的方法育成两用核不育系金山S-1,已于2004年12月通过福建省科技厅技术鉴定。该两用不育系具临界温度低、不育性稳定、不育期长(>120 d)、米质优、配合力好、异交率结实高等特点。以金山S-1配制的杂交稻新组合具产量高、米质优等特点,具有较广阔的应用前景。

光敏核不育水稻育性转换的温度敏感期研究

在14.5小时长光照下,利用人工气候箱对农垦58S和W6154S于幼穗分化不同时期分别进行不同温度处理。育性结果表明农垦58S的育性转换温度敏感期为颖花原基分化期至单核花粉期,W6154S的温敏期为雌雄蕊形成期至单核花粉期。农垦58S的温敏期较长,育性受低温影响后在高温下要较长时期才能恢复不育;W6154S育性易受低温影响,但温敏期较短,在高温下不育性恢复快。在光敏核不育水稻育性感温性鉴定中,建议在雌雄蕊形成期至减数分裂末期进行温度处理。

分子标记辅助选择改良水稻光温敏核不育系华201S的白叶枯病抗性

以华201S为受体亲本,以含Xa21和Xa7基因的抗白叶枯病材料华恢20为供体亲本,进行1次杂交、3次回交和3次以上自交,每个世代用基因内标记PTA248对Xa21进行分子检测,用连锁标记STSP3(与Xa7的遗传距离为0.9 cM)对Xa7进行检测。新选育的4个株系YR7009(Xa21)、YR7014(Xa21)、YR7016(Xa7)和YR7023(Xa7/Xa21)在生育期、主要农艺性状、育性转换特性方面都与华201S基本相似。YR7009(Xa21)和YR7014(Xa21)对PXO61、PXO99、ZHE173和GD1358等4个菌株具有良好的抗性,而对FuJ和YN24不具有抗性。YR7016(Xa7)和YR7023(Xa7/Xa21)对6个菌株都表现良好的抗性,尤其是聚合双基因的YR7023(Xa7/Xa21)对6个菌株都表现高抗。表明已经将广谱白叶枯病抗性基因Xa21和Xa7渗入到华201S中,成功地改良了华201S的白叶枯病抗性。

1Dx5亚基特异PCR标记在小麦品质性状群体改良中的应用

利用1Dx5亚基特异PCR标记对改良品质性状的Ta1小麦轮回选择群体C4进行了分子检测,并通过SDS-PAGE电泳对其结果的验证,以探讨群体5亚基基因型的组成及其特异PCR标记用于轮回选择的可行性。结果表明(1)1Dx5亚基特异PCR标记在具有1Dx5亚基的单株中均扩增出450bp的基因片段,生化标记检测也证明所有能扩增出450bp片段的单株均含有5亚基的条带,分子标记与生化标记结果一致,而且PCR扩增结果的重复性好。说明1Dx5亚基特异PCR标记的选择准确率高,完全可用于Ta1小麦群体改良中1Dx5亚基基因型的检测;(2)构建的轮回选择群体C4中携带1Dx5亚基的优质基因型比例高,达56.81%,且大都伴随与之连锁的10亚基出现。生化检测表明群体中同时产生5+12稀有亚基和14+15与5+10的聚合体。

光敏核不育水稻两个光周期反应的遗传研究

采用湖北光敏核不育水稻第一光周期反应钝感的品种与第一光周期反应敏感的光敏核不育系杂交,F_2代分单株在人工光照条件下进行两个光周期反应的鉴定。结果表明,所用组合的第一光周期反应属数量性状,F_2呈连续单峰曲线分布。第二光周期反应属一对主效隐性基因控制。两个光周期反应分属不同基因(组)控制,但第二光周期反应的不育基因与第一光周期反应感光性强的基因组表现有连锁关系。两个光周期反应的基因(组)可以分离重组,形成第一光周期钝感,第二光周期敏感的光敏核不育系。