利用分子标记分析22种水稻10个抗稻瘟病基因的基因型

为了明确已克隆的部分稻瘟病抗性基因在上海市主栽水稻品种以及新培育的水稻品系中的分布,选取22份水稻为材料,利用分子标记检测技术,对10个已被克隆的抗稻瘟病基因进行基因型鉴定及分析.结果表明,22种水稻均含Pi36、Pi37、Pi-d2、Pib抗性基因,而Pi-kh、Pi9、Pi2、Pi-ta、Pikm抗性基因在22种水稻中分布各有不同,所有被测水稻中均不携带Pi25抗性基因.     

利用分子标记辅助选育优质香软水稻新品系“上师大22号”及试种

为培育适合上海市及其周边地区种植的优质香软水稻，以优质水稻“银香38”为母本，以抗稻瘟病水稻品系“13-2”为父本，进行杂交获得了F1种子；再以抗倒性较强的“武运29185”香稻为母本，与F1植株进行复交，然后多代自交，期间利用香味基因和抗稻瘟病基因分子标记辅助筛选，成功培育出优质香软水稻“上师大22号”.试种结果显示：“上师大22号”水稻全生育期比目前上海市主栽常规水稻“秀水134”短12 d，接近“银香38”水稻；“上师大22号”亩产量分别比“秀水134”和“银香38”增加2.84%和3.96%；“上师大22号”稻瘟病抗性及抗倒性都比“银香38”明显提高.“上师大22号”水稻新品系的成功培育，为上海市进一步推广优质稻品种奠定了基础.     

水稻光敏核不育基因分子标记的研究

石明松（１９７３）从晚粳品种农垦５８大田中发现了具有光周期敏感核不育性状的水稻农垦５８Ｓ,并被正式命名为湖北光周期敏感核不育水稻（ＨＰＧＭＲ）,它的发现及利用可以将杂交水稻种子的生产方式由“三系法”改变为更经济的“二系法”,这将对杂交水稻的生产起到十...     

分子标记辅助选育香型软米水稻“上师大18号”

以香型软米水稻"青香软粳"作为软米基因供体亲本,以香型非软米水稻"光明粳2号"作为转育亲本,结合软米基因分子标记辅助筛选,成功培育出香型软米水稻"上师大18号"."上师大18号"水稻全生育期约148 d,早于"青香软粳"水稻约5 d."上师大18号"水稻主要农艺性状和产量性状都与"光明粳2号"接近."上师大18号"稻米直链淀粉质量分数为8. 2%,符合软米特征."上师大18号"水稻的成功选育有助于上海地区推广生育期相对较短的香型软米水稻.     

上海市11种常规粳稻抗稻瘟病基因分子标记检测

利用分子标记检测技术,对9种参加2020年上海市水稻区域试验的品种和2种本课题组新培育的新品系的共10个抗稻瘟病基因位点进行了检测.结果显示,Pi37,Pi41,Pi-d23个基因在11种水稻中出现的频率达100％,Pi2,Pi5,Pi9,Pi36,Pikm和Pib抗性基因在11种水稻中出现的频率分别为18.18％,9...     

分子标记辅助选育红米巨胚水稻

以正常胚红米水稻"上师大6号"和白米巨胚水稻"上师大5号"作为亲本进行杂交和回交,结合分子标记辅助育种,成功选育出红米巨胚水稻新品系"上师大10号"."上师大10号"的单株重与"上师大5号"差异不显著,但不同地区试种比较结果显示,"上师大10号"产量高于"上师大5号",且生育期较"上师大5号"短4~5 d."上师大10号"的胚重与糙米重比值显著小于"上师大5号",但其胚体积与糙米体积比和"上师大5号"间没有显著差异."上师大10号"的成功培育为市场进一步开发应用红米巨胚水稻提供了物质基础.     

抗稻瘟病和纹枯病的转基因水稻新品系

将水稻碱性几丁质酶基因（RC24）导入优良灿稻品种竹籼B,外源RC24基因可以稳定整合到RO代至R6代转基因水稻基因组中,并得到表达,已获得同时抗稻瘟病和纹枯病的转基因品系竹转68和竹转70以及43个转基因纯合株系。     

水稻温敏核不育系HD9802S不育基因的SSR标记定位

使用分子标记辅助选择可以获得目标性状的个体从而提高育种效率.以HD9802S/(HD9802S/R287) BC1群体为材料,利用BC1群体中随机选择的32个高度不育单株和32个高度可育单株,以及涵盖水稻12条染色体的22个SSR标记,通过连锁分析进行染色体定位,将温敏核不育基因(HDtms)定位于第2号染色体上;利用软件QTL Ici Mapping 3. 0进行染色体分群,HDtms被划分到第2号染色体上.在第2号染色体上覆盖筛选得到10个SSR标记,以回交群体中428个单株为材料绘制连锁图谱,图谱全长112. 21 cM,平均图距为11. 21 cM,HDtms在标记RM5897与RM12783之间.在此区间内筛选到与HDtms连锁紧密的SSR标记.其中RM12747是得到筛选率最高的分子标记,为90. 135%,且在HD9802S与R287间具有多态性.说明对温敏不育性的分子标记辅助选择可初步使用该分子标记.     

耐温牙鲆分子标记辅助选育研究

应用分子标记进行耐温牙鲆的辅助选育.首先应用已知的与耐温性为极显著负相关的牙鲆微卫星引物Po42对50尾牙鲆亲鱼DNA进行PCR分析,根据PCR分析结果将其分成耐温组和非耐温组.经过人工催产,耐温组的产卵量明显大于非耐温组和对照组.耐温组繁育的子代仔鱼变态率明显比非耐温组和对照组子代高.在人工增温的条件下,耐温组仔鱼成活率比非耐温组和对照组的仔鱼显著提高.本研究结果证明,牙鲆微卫星引物Po42可用于耐温牙鲆的分子标记辅助选育,同时也进一步确认其与牙鲆耐温性状具有关联性.     

分子标记辅助选育香型巨胚水稻

香型稻米因其在蒸煮和食用时都会有特殊的香味受到人们的喜爱．研究中,分别以实验室培育的香型正常胚水稻和非香巨胚水稻“上师大5号”作为亲本进行杂交,结合水稻香味基因分子标记辅助常规育种成功选育出香型巨胚水稻“上师大8号”．比较分析“上师大5号”和“上师大8号”两种巨胚水稻主要农艺性状和产量性状．结果显示：虽然“上师大5号”水稻平均每穗实粒数极显著地超过“上师大8号”水稻,但是“上师大8号”有效穗数略多于“上师大5号”水稻,而且千粒重也略高于“上师大5号”水稻,由此两者平均单株重接近,分别为29．10g和28．92g,统计分析差异不显著．两种巨胚水稻胚性状分析显示：“上师大8号”巨胚水稻的胚重量以及胚体积都与“上师大5号”巨胚水稻统计分析无显著差异,而且两种巨胚水稻的胚与糙米重量比以及胚与糙米体积比,统计分析都没有显著差异．开展此研究,为今后香型巨胚水稻的市场开发应用奠定了重要基础．     

Analysis of rice blast resistance genes by QTL mapping

Resistance to rice blast pathogen mostly shows a quantitative trait controlled by several minor genes. Its complexity and the mutable characteristic of rice blast isolates both hinder the development of the blast resistance research. The article here tried to explore the resistance gene distribution on rice chromosomes and the way of function. Totally 124 QTLs have been identified against 20 isolates using Cartographer software with a ZYQ8/JX17 DH population, which separately are at 100 loci of 72 marker intervals on 12 rice chromosomes. Of them, 16 QTLs were determined by the isolate HB-97-36-1. 82 QTLs (66.13%) are from the resistant parent alleles, ZYQ8, while 42 QTLs (33.87%) are from the susceptible parent alleles, JX17. In comparison of their positions on chromosome, most QTLs are clustered together and distributed nearby the major genes especially the regions on chromosomes 1, 2, 8, 10 and 12. Each QTL could account for the resistance variation between 3~2%-68.64%. And, a positional QTL might display the resistance to several different isolates with different contributions.     

水稻光敏核不育性的遗传及相关分析

以花粉深染率、自然结实率为育性指标 ,于 2 0 0 1年观察分析了农垦 5 8S× 1 5 1 4F2 群体植株在武昌 (3 0°3 0′N)自然长日照条件下的育性表现 .实验结果表明农垦 5 8S的光敏不育性受两对核内隐性主基因控制 ,并受微效基因的修饰作用 .另外 ,对有关性状的相关分析结果表明 ,株高与抽穗期、花粉深染率和自然结实率呈现显著相关 ,花粉深染率或自然结实率与抽穗期不相关 ,花粉深染率与自然结实率呈高度正相关 (r=0 .75 74) .     

Restorable variation of cytoplasmic type of male sterile line in rice by in vitro culture

Rice male sterile (MS) lines, IR66707A and IR69700A, which possess the cytoplasm of Oryza perennis and O. glumaepatula respectively, belong to the cytoplasmic type. Their sterility could be maintained but not be restored. By somatic cell culture of these two MS lines, 47 somaclones with 465 R1 plants were obtained. All of the 465 R1 plants were sterile in the spring season in Guangzhou. According to the expression of the R1 plants and the level of similarity to their donor parents, they could be divided into three types. The plants of type Ⅰ were male sterile. The sterility of some somaclones of this type could be restored by the test crossing varieties or alternated to fertile by changes of some environmental conditions. The hybrid F1 of test cross from the MS somaclones in type Ⅰ was fertile while the hybrid F1 from the donor MS lines was still sterile. The R1 plants of type Ⅱ were similar to the donor parents and also male sterile. The hybrid F1 from all of the plants of type Ⅱ crossed to test variety were still sterile, so they did not possess restorability. For the somaclone of type Ⅲ, all of R1 plants were sterile in both male and female organs. No seed was set in both conditions of self and cross pollination. The fact that the restorable variants obtained in the cytoplasmic type of MS lines of rice by in vitro culture reported here should be the first sample in somaclonal variation in plant kingdom.     

Molecular cloning, chromosomal mapping and expression analysis of disease resistance gene homologues in rice (Oryza sativa L.)

Resistance-like sequences have been amplified from first strand cDNA and genomic DNA of rice by PCR using oligonucleotide primers designed from sequence motifs conserved between resistance genes of tobacco andArabidopsis thaliana. 3 PCR clones, designatedOsr1, Osr2 andOsr3 which were 98% identical in nucleotide sequence level, have been found to be significantly homologous to known plant resistance genes and all contained the conserved motifs of NBS-LRR type resistance genes, such as P-loop, kinase2a, kinase3a and transmembrane domain.Southern hybridization revealed that rice resistance gene hornologueswere organized as a cluster in the genome. RFLP mapping using a DH population derived from anindica/japonka cross (Zhaiyeqing 8/Jingxi 17) and an RFLP linkage map assigned two copies ofOsrl and one copy ofOsr3 to the distal position of chromosome 12 where a blast resistance QTL has been mapped previously. Northern blot analysis showed thatOsrl gene was constitutively transcribed in rice leaves, shoots and roots. Further study concerning isolation of full-length cDNAs would be conducive to elucidating the functions of these genes.     

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.     

Identification of differentially expressed genes in photo-period sensitive genie 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.