RAPD在玉米温敏型核雄性不育研究中的应用

对适用于玉米的ＲＡＰＤ实验体系进行研究,摸索出适用于玉米的ＲＡＰＤ的反应适宜条件闻稳定的玉米ＲＡＰＤ分析体系,应用这一体系对温敏核雄性不育玉米琼４２－Ｑｍｓ及其可育的近等基因系琼４２进行ＲＡＰＤ分析,在３００多个引物中发现１个引物在两种中扩增带型有差异,经重复试验,初步认为此差异与玉米雄性不育基因连锁。     

用人工冷水池鉴定水稻光温敏核不育系雄性不育起点温度的研究

用人工冷水池与人工气候箱分别对8个水稻光温敏核不育系在它们育志换温敏感期进行5d日均温23.5℃（22－25℃）的冷水与6d日均温23．5℃（19．5－26．0℃）的冷气处理。结果表明：株S－1、陆18S、96－5－2S、139S及康201S在两种低温处理条件下均表现不育，自交结实率与花粉可染率均为0；培矮64S在人工冷气处理条件下也表现不育，自交结实率与花粉可染率为0，但在人工冷水处理条件下表现轻度可育，自交结实率为0．05％，花粉可染率为5．0％；810S与179S在两种低温处理条件下均表现可育，自交结实率为3．7％－18．5％，花粉可染率为4．3％－41．0％，但人工冷水处理条件下的自交结实率与花粉可染率明显高于人工冷气处理条件下的自交结实率与花粉可染率。说明用人工冷水池鉴定水稻光温敏核不育系雄性不育的起点温度是可行的，而且鉴定的灵敏度比人工气候箱更高。根据人工冷水池与人工气候箱鉴定的结果可知：株S－1、陆18S、96－5－2S、139S及康201S的雄性不育起点温度小于23．5℃，培矮64S的雄性不育起点温度在23．5℃左右，而810S与179S的雄性不育起点温度大于23．5℃。     

棉花核雄性不育系杂交成铃率的影响因素研究

研究了双隐性核雄性不育系在不同气象条件下、不同时间人工辅助授粉配制杂交组合的杂交成铃率以及不同品种 (系 )作父本的杂交成铃率。结果表明 :以核雄性不育系为母本的杂交组合成铃率与气温、日照时数呈显著正相关 ,与降水量呈显著负相关 ;7月 2 0日以前与 8月 2 0日以后的杂交成铃率较高 ,每天 9- 1 1时的杂交成铃率高于 1 4- 1 6时 ;常规品种 (系 )与国产抗虫棉作父本的杂交成铃率较高 ,美国抗虫棉作父本的杂交成铃率较低 ,海岛棉作父本的杂交成铃率高但单铃种子数少。用核雄性不育系为母本配制杂交组合最佳授粉的气象条件为平均气温在 30℃左右、最高气温不超过37℃、日照长、无降水 ;最佳授粉时间在 8月 2 5日以前的每天 9- 1 1时 ;宜选择常规品种 (系 )或国产抗虫棉作父本     

茉莉酸刺激的橡胶树胶乳cDNA消减文库的构建及其序列分析

采用抑制性消减杂交(suppression subtractive hybridization,SSH)技术,构建了外源茉莉酸刺激条件下橡胶树胶乳与未处理橡胶树胶乳差异表达的cDNA消减文库,经蓝白斑筛选共得到121个含有插入片段的阳性克隆。通过菌落PCR的方法分别对阳性克隆的插入片段进行扩增,结果表明,95％左右阳性克隆插入片段的大小在200-600 bp之间。随机选取25个克隆进行测序,并对所得的25条表达序列标签(EST)序列用Blastn(基本局域联配搜寻工具)检索基因文库(genbank),其中10条EST片断可找到碱基序列相似性大于80％以上的同源基因序列,其余15条EST为没有任何功能线索的未知序列。外源茉莉酸刺激条件下橡胶树胶乳cDNA消减文库的构建和在此基础上克隆橡胶树胶乳中JA信号的候选应激基因,将为开展茉莉酸调控橡胶树胶乳代谢和橡胶生物合成的分子机理研究打下基础。     

核盘菌诱导下甘蓝型油菜宁RS-1 的SSH文库构建

以甘蓝型油菜宁RS-1为材料,用核盘菌(Sclerotinia sclerotiorum (Lib.) de Bary)对6叶期幼苗进行接种。提取经核盘菌诱导前和诱导后的宁RS-1叶片mRNA,经反转录成cDNA,以及经过酶解后加接头和两次杂交,构建差异表达的抑制消减(SSH)cDNA文库。正向消减文库是以诱导后的宁RS-1 cDNA为tester,以正常的宁RS-1 cDNA为driver。所构建的正向文库重组率达到96.2%,反向文库重组率达到94.5%,文库容量均达到了2 190和2 780,表明所构建的文库质量良好。对从文库中随机挑取的克隆进行测序分析,获得了与抗病和代谢有关的基因,如γ-谷氨酰半胱氨酸酶基因、抗真菌蛋白基因、铁蛋白基因、ACC氧化酶基因等,为开展油菜抗菌核病重要功能基因的克隆和鉴定奠定了基础。     

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...     

水稻光温敏核雄性不育系培矮64S 花粉败育的细胞学机理

 采用半薄切片及薄切片技术, 对水稻光温敏核雄性不育系培矮64S 与正常水稻品种IR36 的花粉形成发育过程进行了比较研究。结果表明, 小孢子母细胞减数分裂之前, 培矮64S 与IR36 的发育过程基本相似。但从减数分裂开始, 培矮64S 的雄性性细胞出现一些异常变化, 并最终发育至二胞花粉早期败育。这些变化主要发生在两个阶段: (1) 减数分裂前期,约半数的小孢子母细胞的胞质出现异常, 游离核糖体稀少, 并具有不发育线粒体和大量泡状内质网。这类异常的小孢子母细胞在随后的发育中逐渐液泡化并最终解体。(2) 早期小孢子形成之后, 几乎所有小孢子外壁均发育异常, 表层与里层之间界限不清, 缺少中间透明带, 同时内壁没有形成。但是, 在花粉发育的整个过程中, 培矮64S 与IR36 绒毡层的发育和降解过程基本相似。据此认为, 培矮64S 的花粉败育可能是由小孢子母细胞或花粉外壁的异常发育造成的, 而不是由绒毡层发育引起的。     

非环境敏感型核雄性不育水稻育性的化学调控初报

非环境敏感型核雄性不育水稻育性的化学调控初报徐仁胜（中国水稻研究所植物生理系．杭州３１０００６）关键词：核雄性不育;育性恢复;植物生长调节剂ＣｈｅｍｉｃａｌＲｅｓｔｏｒａｔｉｏｎｏｆＦｅｒｔｉｌｉｔｙｏｆＧｅｎｉｅＭａｌｅ－ｓｔｅｒｉｌｅＲｉｃｅｗｉ...     

大豆突变体NJS-1H核雄性不育性的细胞学与遗传学分析

利用化学诱变剂叠氮化钠(NaN3)处理大豆品种南农86-4,获得雄性不育突变体NJS-1H。细胞学观察发现,该突变体不育株NJS-1H(s)的小孢子在减数分裂前期I染色体联会异常,出现单价体;减数分裂过程中出现染色体落后、不对称或不同步分裂等现象;在四分体阶段,出现各种畸形多分体及大量微核;所形成单核小孢子细胞核消失,细胞质变稀薄,最后成熟花粉粒无内容物,完全不育。从减数分裂到花粉粒发育成熟都有败育发生,但大量败育主要发生在减数分裂阶段。人工平行杂交试验表明其雌性育性不正常。对突变体后代育性分离株系及株系群的遗传分析,发现NJS-1H的雄性不育性受1对隐性核基因控制。     

爪哇稻Wanilava细胞质雄性不育系的创建及AFLP指纹图谱分析

初步鉴定了爪哇稻Wanilava细胞质雄性不育系的特征特性,并以2套水稻（Oryza sativa）同核异质材料273JW（爪哇型胞质不育系）、273D（D型胞质不育系）及其对应的保持系273B, 803JW（爪哇型胞质不育系）、803WA（野败型胞质不育系）及其对应的保持系803B为研究对象,利用AFLP 分子标记技术构建了JW型不育胞质的指纹图谱,分离了JW型胞质与野败型胞质及保持系的差异片段。     

温敏核不育水稻植株温度的变化规律及与环境关系的模型

以温敏核不育水稻培矮64S为材料,采用10～15 cm水层灌溉处理和无水层对照,对植株温度及其与植株冠层小气候和灌溉水因子的关系作了分析。水稻植株温度与150 cm大气温度在数值和相位上均存在一定差异。8:00～20:00植株温度均明显低于大气温度,21:00～次日7:00植株温度与大气温度基本相同;日最高植株温度出现在13:00,比最高大气温度提前1 h,但日最低植株温度和最低大气温度均出现在6:00;植株温度的平均日较差比气温小。在同一高度上相比,晴天6:00～13:00植株温度比空气温度高,而且提前1 h升温,18:00～次日6:00则两者逐渐趋同或植株温度稍低;而在阴天,植株温度则全天一直高于空气温度,最高温度出现的时间也相同。植株温度白天的变化主要受太阳辐射的影响,天空状况（云量或日照时数）和风速都通过对辐射强度的调节和热量的交换而产生作用。植株温度夜间的变化主要受灌溉水的影响。在本试验条件下,日平均气温（Ta）29.6℃是灌溉水提高或降低植株温度的临界温度值,当Ta＞29.6℃时,灌溉水具有降低植株温度的作用,反之,灌溉水具有提高植株温度的作用。植株温度与水 气温差符合二次曲线关系。植株冠层在白天吸收或反射太阳辐射,夜间则阻挡热量散失,对调节植株温度具有明显的缓冲效应。通过相关分析和回归拟合,建立了两个可供实用的水稻植株温度的环境模型。     

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.     

水稻光温敏核不育机理设想及光温敏核不育系选育策略

为更深入了解水稻光温敏核不育的遗传机理,综述了我国水稻光温敏核不育机理研究取得的成就及存在问题,提出了水稻光温敏核不育机理新设想,即水稻光温敏不育系中不存在光敏不育基因和温敏不育基因,其育性转换是主效不育基因与发育感光基因或(和)发育感温基因相互作用的结果;正常水稻品种中存在的不育基因位点(微效不育基因)可影响光温敏核不育系的不育起点温度,微效不育基因聚合越多,则不育系不育起点温度越低;微效不育基因完全纯合,则不育起点温度不会漂变。基于不育机理新设想,提出了光温敏核不育系选育的策略,即不同生态区、不同生态类型的光温敏核不育系选育的光温指标不一样,增压选择是选育低不育起点温度核不育系的技术核心,全面提高综合性状水平和配合力是选育实用光温敏核不育系技术的关键。     

湖南新选育的水稻光温敏核不育材料的育性研究

对湖南省近期内新选育的水稻光温敏核不育材料的育性进行了比较研究。在恒温２４℃条件下，有７个材料的花粉可染率和结实率均为零；４个材料有极少量可染花粉，结实率为零。在模拟１９８９年盛夏低温条件下，有６个材料的花粉可染率和结实率均为零；有６个材料有极少量可染花粉，结实率为零。     

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.     

隐性长穗颈温敏核不育水稻长选3S穗颈节间伸长规律的研究

以隐性长穗颈温敏核不育水稻长选3S及其亲本培矮64S为材料,研究了温度对长选3S穗颈伸长的影响。人工24℃条件下,在始花前第12天至始花前第4天和始花当天至始花后第3天两个时段,穗颈节间伸长速度慢,但从始花前第4天至始花当天,长选3S穗颈节间伸长速度最快,其节间日均伸长的长度是培矮64S的2.1倍。在敏感期分别进行22℃、24℃、26℃和28℃ 4种人工温度处理, 28℃条件下穗颈节间伸长受阻, 出现包颈现象;22~26℃条件下穗颈伸出长度都为正值,不包颈,且穗颈伸出剑叶叶鞘的长度随温度降低而增加。对穗颈节间细胞数目和细胞长度的比较分析表明,长选3S穗颈节间的伸长主要是始花前第12天至始花期节间细胞分裂和细胞伸长共同作用所致,其中后者作用更为显著,且随处理温度的升高,穗颈节间最内和最外层薄壁细胞数目减少,细胞平均长度下降。     

温敏核不育基因在籼型三系遗传背景下的育性表达

用3个温敏核不育系培矮64S、6311S和360S与7个籼型质核互作型水稻雄性不育系及相应的保持系、3个恢复系配组,观察了51个组合的F1、19个F2及6个BC1的育性表现。结果表明：培矮64S温敏核不育性状由2对隐性基因控制,具有对质核互作型雄性不育系育性的强恢复基因;6311S温敏核不育性状由1对隐性基因控制,同时具有1对弱恢复基因;360S温敏核不育性状由1对隐性基因控制,但没有恢复基因。进一步利用4个细胞质雄性不育系/6311S组合F2群体中4个温敏核不育株与5个细胞质雄性不育系CMS配组,研究了杂交F1的育性,表明在可育细胞质背景下,三系恢复基因对温敏核不育基因的表达没有影响;在不育细胞质背景下,三系恢复基因是温敏核不育基因表达的关键。由此提出了选育不育细胞质背景的光温敏核不育系和温敏核不育背景的质核互作型不育系的策略。     

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.     

利用cDNA—AFL吩析辣椒细胞核雄性不育两用系的基因表达差异

[目的]为进一步了解辣椒不育性的分子遗传机理和雄配子的发育机制,对辣椒细胞核雄性不育的育性相关基因进行初步研究。[方法]利用eDNA—AFLP对辣椒细胞核雄性不育两用系‘AB23’的可育株花蕾和不育株花蕾的表达基因进行比较和分析。[结果]可育株与不育株在基因表达上存在差异,筛选出10对引物,并得到了有差异且重复性好且的19条TDFs,其中14条TDFs功能已知,3个功能未知,2个未找到同源序列。该研究通过RT—PCR技术对获得的19个TDFs进行验证,其表达特征与eDNA—AFLP结果一致。[结论]差异基因功能涉及细胞分泌、代谢、转录、细胞程序性死亡、蛋白质修饰、胞间运输、信号转导、细胞黏附等多个相关过程。     

Marker-Assisted Breeding of Thermo-Sensitive Genic Male Sterile Line 1892S for Disease Resistance and Submergence Tolerance

Rice line 1892S is an elite thermo-sensitive genic male sterile(TGMS)line for two-line hybrid rice production.However,1892S is susceptible to rice blast,bacterial blight and submergence.Here we reported the introduction of blast resistance(R)gene Pi9,bacterial blight R gene Xa21 and submergence tolerance gene Sub1A into 1892S genetic background through backcrossing and marker-assisted selection.The improved TGMS line 31892S and its hybrids conferred disease resistance to rice blast and bacterial blight,and showed submergence tolerance for over 14 d without significant loss of viability.The sterility-fertility conversion of 31892S was similar to that of 1892S.31892S and its derived hybrid rice had similar agronomic traits and grain quality with 1892S and the control hybrid rice,respectively.The newly developed 31892S provided an improved TGMS line for two-line hybrid rice production with disease resistance to rice blast and bacterial blight,and submergence tolerance with no yield penalty or change in grain quality.