Development and Application of SCAR Markers for Discriminating Cytoplasmic Male Sterile Lines from Their Cognate Maintainer Lines in Indica Rice

The DNAfragments about 1 600 bp were amplified using random amplified polymorphism DNA （RAPD） primer OPAl2 with the templates of mitochondrial DNA of Zhenshan 97A and Zhenshan 97B, and were sequenced. The nucleotide sequences and lengths of the fragments from Zhenshan 97A and Zhenshan 97B showed no difference. The precise length of the fragment was 1 588 bp. Sequence characterized amplification region （SCAR） primers were then developed to discriminate the cytoplasmic male sterile （CMS） lines and their maintainer lines. A specific 1 588 bp fragment could be amplified with SCAR primers, CHI19F2/CHI19R2 and CHI20F3/CHI23R3, in the mitochondrial DNA of Zhenshan 97A, but not Zhenshan 97B. Furthermore, the specific fragment could be also amplified from the total DNA from green leaf tissues of Zhenshan 97A with SCAR primers, but not Zhenshan 97B. With the corresponding primers, the specific fragment could also be amplified from the total DNA of green leaves of other two CMS lines with wild abortive type cytoplasm （CMS-WA）, namely Zhenpin A and Tianfeng A, but not in their maintainer lines. Moreover, using total DNA as template, each of the four pairs of SCAR primers could also be used to amplify the 1 588 bp fragment in CMS-ID （Indonesia paddy type） line 11-32A but not in 11-32B, and the specific fragment was amplified from the DNA of both F1 and F2 seedlings of Shanyou 63. The results of detecting the genetic purity of a man-made mixture of the seeds of Zhenshan 97A using CHI20F3/CHI23R3 were completely consistent with the phenotypes. Taken together, these results indicated that the specific 1 588 bp-fragment amplified by CHI20F3/CHI23R3 was the unique amplification products of CMS mitochondrial DNA, and could be used to distinguish CMS-WA and CMS-ID lines from their corresponding maintainer lines at the seedling stage.

Comparisons on Genetic Diversity among the Isonuclear-Alloplasmic Male Sterile Lines and Their Maintainer Lines in Rice

Four sets of rice isonuclear-alloplasmic lines including 16 male sterile lines and their maintainer lines were analyzed by using 91 pairs of SSR primers to study the genetic diversity of nuclear genome and their relative relationships. A total of 169 alleles were detected in the 16 lines, with a frequency of polymorphic loci of 53.85% and an average number of alleles per locus of 1.8, and the average gene diversity was 0.228. Four sets of the isonuclear-alloplasmic male sterile lines shared 146 identical alleles, corresponding to 86.39% of the total alleles; meanwhile, there are 23 different alleles among the tested materials, being 13.61% of the total alleles. On average, 78.70% identical alleles and 21.30% different alleles of the total alleles were detected between the isonuclear-alloplasmic male sterile lines and their maintainer lines. There were 53.85% identical alleles and 46.15% different alleles of the total alleles among the homozygous allonucleus male sterile lines. The fingerprints were established for some male sterile lines and maintainer lines. All the materials tested were divided into three groups at the 0.2 genetic distance based on the cluster analysis. Eight lines of Huanong A and Huayu A （including Huanong B and Huayu B） were in the first group, four lines of Kezhen A （including Kezhen B） in the second group, and four lines of Zhenshan 97A （including Zhenshan 97B） in the third group. For the isonuclear-alloplasmic male sterile lines, the similarity coefficient between Y （Yegong） type and WA （wild abortive） type or between CW （Raoping wild rice） and WA type reached 87-98%.

Breeding of Maintainer Line and Sterile Line Brown Planthopper Resistant Rice Using Marker-assisted Selection

Guangxi common wild rice variety brown planthopper highly resistant introgression line HS204 was taken as antigen donor material,hybridization,backcrossing,and molecular marker-assisted selection methods were adopted,to select maintainer line and sterile line materials,so as to provide excellent material foundation for resistance breeding of hybrid rice. Through the marker-assisted selection,it obtained 4 pieces of homozygous resistance gene maintainer line intermediate materials( 100B,101B,102B and 103B),and 2 pieces of resistant sterile line materials( 100A and 103A). All 10 combinations that have testcross with highly resistant sterile 100A showed higher level of brown planthopper resistance: 100A/R2586,100A/KR838,and 100A/KR527 had high resistance level,the others had low to intermediate resistance;100A/KR527,100A/R2586,100A/Minghui 63,100A/Fuhui 838 and 100A/Gui 99 combinations had yield per plant significantly higher than the control group( Teyou 7118),increasing by 14. 45%-49. 26%. The obtained resistant lines are expected to provide a better gene source for the breeding of resistant sterile lines of hybrid rice and the obtained resistant sterile lines can be directly used in the selection of three-line hybrid rice.

Thermal Resistance of Common Rice Maintainer and Restorer Lines to High Temperature During Flowering and Early Grain Filling Stages

Fifteen common rice maintainer lines and 26 high-yielding restorer lines were used to evaluate their thermal resistance and fertility during flowering and early grain filling stages. The rice plants were subjected to high temperature stress （39-43 ℃） for 1-15 d from main stem flowering. Based on the heat stress index, they were divided into thermal resistant lines, semi-thermal resistant lines, semi-thermal sensitive lines and thermal sensitive lines. Therefore, the maintainer lines K22B, Bobai B and V20B belonged to thermal resistant lines, whereas 11-32B, Zhongzhe B and Zhong 9B belonged to thermal sensitive lines. For rice restorer lines, Minghui 63 had the highest thermal resistance, followed by R207, P32, P929, and the lowest thermal resistant lines P62-2-2, R8006 and P51. The correlation analysis indicated that the heat stress index was significantly correlated with seed-setting rate and abortive grain rate under heat stress, but not under natural conditions. This indicated that heat stress occurred during flowering and early grain filling stages mainly decreased the seed- setting rate and significantly increased the abortive grain rate in both rice maintainer and restorer lines.

Resistance Evaluation of Some Chinese Leading Rice Maintainer, Restorer lines and Their Hybrids to Magnaporthe grisea

Six isolates of Magnaporthe grisea were selected to inoculate on 10 Chinese leading maintainer lines (B-lines), 14 restorer lines (R-lines) and their F1 hybrid plants. In the tested rice materials, R-lines were proved to be more resistant to blast than B-lines. The resistance frequency of about 25% F1 hybrid plants was less than their parents. In addition, 26 isolates of M. grisea collected from different rice growing areas of China were inoculated on 13 new improved hybrid rice combinations. The resistance frequencies of 5 improved hybrids were better than those of the controls and leading varieties in rice production of China.

Difference and Inheritance of the Sterility Maintenance of CMS in Indica Rice

The inheritance of the sterility maintenance ability was studied for three types of CMS lines (Zhenshan97A, XieqingzaoA and II-32A) and several lines in indica rice. The result showed that the abilities to keep the male sterility of the maintainer lines strengthened obviously with the progressive backcrossing. The critical stage to maintain the sterility of CMS lines happened usually in BC5-6 generation. The selection of the recurrent parent gave an obvious influence on the pollen fertility distribution of the later generations. The author has discussed ways of improving of the maintainer lines.

Development and verification of the cytoplasmic-nuclear male sterile soybean line NJCMS1A and its maintainer NJCMS1B

THE first cytoplasmic male sterile soybean line was reported by Davis in a U.S. Patent, but no further information has been released since then. Sun et al. developed a cytoplasmic-nuclear male sterile soy-