Preliminary Studies on the Effect of Photoperiod and Temperature on Male Fertility of a Cytoplasmic Male Sterile Line in Wheat

[Objective] The study aimed to reveal the effect of photoperiod and temperature on male fertility of cytoplasmic male sterile line Vtai911289a in wheat and discuss the mechanism of male fertility alteration. [ Method ] The sowing-date tests and designed conditions were conducted during 2003 -2005. [ Result] Fertility of Vtai911289a, could alter under specific photoperiod and temperature conditions. Temperature is one of the main factors influencing male fertility of the male sterile lines. Vtai911289a, showed stable sterility under the condition of the mean of daily temperature at fertility sensitive stage lower than 19℃ and presented partial fertility when the mean of daily temperature at fertility sensitive stage lower than 20 - 22℃. Photoperiod to some extent affects the male fertility of Vtai911289a, long-day condition is helpful for the male fertility of the sterile line. [ Conclusion] The application of photoperiod temperature-sensitive cytoplasmic male sterile line in production has a higher safety than that of temperature sensitive sterile line.

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.

Influence of Genetic Drift of Restoring Gene (Rf) on Seed Purity of Yuetai A, a Honglian-Type Cytoplasmic Male Sterile Line in Hybrid Rice

The seed samples of Yuetai A, a Honglian （HL） type cytoplasmic male sterile （CMS） line in hybrid rice were investigated to assess the seed purity and to analyze the cause of off-type plants by imitating the biological contaminant to Yuetai A in Nanjing, Jiangsu Province and Lingshui, Hainan Province during 2004-2006. The seed impurity of Yuetai A mainly resulted from the genetic drift of restoring fertility gene （Rt） after biological contamination but not from its sterility unstability. All of the ten maintainer lines, five restorer lines and three thermo-sensitive genic male sterile lines used in the study could pollinate Yuetai A and Yuetai B to produce F1 plants, directly or indirectly resulting in Rf-gene drifting into Yuetai A and generating ＇iso-cytoplasm restoring-generations＇. Furthermore, high outcrossing rate and similar heading date of Yuetai A with many varieties used in rice production might easily result in the biological contamination. After removing all plants with Rf-gene mixed in Yuetai A and preventing Rf-gene drifting into Yuetai A, the seed purity of Yuetai A and Yuetai B had been raised to 100%.

Genetic Mechanism of Dominant Earliness in Kefeng A,a New Rice Cytoplasmic Male Sterile Line

Kefeng A is an early maturing indica cytoplasmic male sterile （CMS） line of rice. Combinations derived from Kefeng A and late maturing indica restorer lines showed dominant earliness to various extents. To understand the genetic basis of dominant earliness, the genotype of photoperiod-sensitive genes in Kefeng A was analyzed using a complete set of heading time near isogenic lines （NILs） EGO to EG7, ER, LR, T65, T65E^b, T65E^bm, T65m, NIL（Hd1） and NIL（Hd4）. Results indicated that Kefeng A contained two dominant photoperiod-sensitive alleles E1 and Se-1^U on E1 and Se-1 loci, respectively, and the genotype of photoperiod-sensitivity genes for heading time in Kefeng A was E1E1e2e2E3E3Se-1^USe-1UEf-1Ef-1. Based on the detected heading time genotype, in combination with the heading time of Kefeng A and the early maturing phenomenon in its derived F1 hybrids, it is speculated that Kefeng A might carry a dominant inhibitor gene Su-E7 for the dominant photoperiod- sensitive gene E1, and a recessive inhibitor gene i-Se-1 for another dominant photoperiod-sensitive gene Se-1. The reason why F1 hybrids from Kefeng A exhibited early maturing was hereby analyzed and the breeding value of dominant earliness related genes in Kefeng A was discussed.

Differences in Nuclear DNA Between Male-Sterile and Male-Fertile Lines of Sorghum bicolor

Cytoplasmic male sterility(cms)is determined by nuclear-cytoplasmic interactions. Up to now, most studies are focused on the comparison of cytoplasmic DNAs of male-sterile lines and male-fertile lines, and analysis of nuclear DNA has not been documented yet. In order to find out the possible difference in nuclear genome of male-sterile line A1 Tx623 and corresponding male-fertile line Tx623 of sorghum, random amplified polymorphic DNA(RAPD)approach was used to analyze their cytoplasmic and nuclear genomes. Total DNAs of them were amplified at first to screen primers, which were able to generate reproducible bands specific to male-sterile line or male-fertile line. Then the selected primers were used to amplify their mitochon-drial DNA(mtDNA)and chloroplast DNA(cpDNA). The origins of all the polymorphic fragments were analyzed. After ruling out those amplified from cytoplasmic DNA, seventeen polymorphic fragments were determined to be amplified from nuclear DNA. These fragments originated from nuclear DNA indicate that differences in sequence exist between the nuclear DNA of male-sterile line and male-fertile line of sorghum, which do not agree with the traditional standpoint that they have identical nucleus.

Maxie A- a novel cytoplasm male sterile line

The pollen abortion of Maxie A and Zhengshan97 A belong to stainless type with Ksolu-tion.The maiority of Maxie A abortive Pollenstook cireular shape and few was staining abortion

Analysis of Heterosis, Combining Ability and Heritability of Cadmium Content in Brown Rice of Three-line Indica Hybrid Rice

Five cytoplasmic male sterile （CMS） lines were used as parents in an incomplete diallet cross and six restorer lines of rice design （Nc II design）. Thirty hybrid combinations with the same growth period were selected as experimental ma- terials to investigate the heterosis, combined ability and heredity of Cd content in brown rice of indica hybrid rice. According to the results, Cd content in brown rice showed a significantly negative heterosis; the general combining ability and specific combination ability of Cd content in CMS and restorer lines both reached extremely significant level （P〈0.01）, indicating that both genetic improvement of parents and e- valuation of combinations are important to the breeding of hybrid combinations with low accumulation of Cd; the broad-sense heritabitity and narrow-sense heritability of Cd content were both relatively high with slight differences, which respectively reached 97.73% and 80.10%, indicating that Cd content in brown rice mainly de- pends on the additive action of genes; in addition, parent improvement showed bet- ter effect on the selection of early generation.

Lodging Resistance of Aromatic CMS Line Zhongzhe A and Its Derived Lines

Nineteen rice combinations derived from Zhongzhe A and its derived lines, as well as Y Liangyou 1 as the control, were used to study lodging resistance by detecting plant height and some of the traits of culm and panicle that are related to lodging. The results indicated that the lodging resistance of 16 combinations was significantly stronger than that of Y Liangyou 1. Moreover, the lodging resistances of all combinations derived from Zhongzhe 2A, Zhongzhe 3A, Huazhe A and Zhong 1A were significantly higher than that of Y Liangyou 1. Additionally, the traits related to lodging were not identical within different CMS rice lines, and the relationship between plant height and lodging resistance was only significantly (P < 0.01) detected in Zhongzhe 2A. Some traits of the second internode were closely related to lodging in all combinations, which can be used as an effective index in lodging resistance breeding in rice.

棉花晋A细胞质雄性不育系小孢子发生的显微和超微结构观察

研究了棉花晋 A细胞质雄性不育系的细胞学特征。结果表明 ,1晋 A不育系雄性败育的主要时期是在造孢细胞增殖——小孢子母细胞形成时期。大部分小孢子母细胞在形成过程中退化解体 ;少数小孢子母细胞能进一步发育并开始减数分裂 ,但都仅停留在减数分裂前期 ,该不育系绒毡层提早解体与小孢子母细胞解体有关。 2小孢子母细胞和绒毡层细胞中的线粒体异常 ,对晋 A不育系小孢子败育有直接的影响。线粒体异常表现的特征为 :线粒体肿胀破裂 ,内嵴溶解。

Ogura细胞质甘蓝雄性不育系选育及其利用

利用Ogura不育源，对上海地区不同类型的甘蓝材料进行转育，获得了两个Ogura细胞质甘蓝雄性不育系94BC－15，94BC－12。这两个不育系的不育性稳定，克服了不育源存在的蜜腺退化及雌蕊畸形造成结实不良的生理缺陷，并基本克服了苗期低温黄化现象，具有利用价值。利用不育系94BC－15与亲本92－B6杂交育成了杂种一代“沪甘一号”，该杂种一代具有生长势强、耐热性好、抗霜霉病和TuMV、综合园艺性状优良等特点，产量比“夏光甘蓝”增产10％左右。

小麦K、V、T、CHA细胞质雄性不育类型的光合特性分析

利用2套同核异质的K、V、T、CHA型不育系及其保持系，对其不同生长发育时期的光合特性及光合产物的积累进行分析研究，探讨不同小麦雄性不育类型的胞质效应。结果表明：（1）叶绿素含量在各个材料中表现不同，K、V、T型不育系与保持系相差不大，也未表现出不良效应，CHA型抽穗期的叶绿素含量为2．368—1．253mg／g，较K、V、T型不育系及其保持系低0．668～1．719mg／g，差异显著，开花期后差距不断缩小，趋于一致；（2）K型不育系的Fv／Fm值大多保持在0．84以上，其他不育系及保持系多在0．77～0．84之间，其中ФPSⅡ也相对较高，而T型胞质类型的Fv／Fm值相对要低，在孕穗期仅为0．77～0．806，后期有所增加；（3）保持系从抽穗期到灌浆后期Pn值多在10以上，而K、V、T型3种不育胞质类型表现出负效应，尤其在灌浆后期差异显著。在抽穗期，K型不育系的Pn值较低，T型不育系的Pn值下降幅度最大，在灌浆后期降到1．45～1．9，显著低于K、V型的1．55～2．85，太911289CHA型不育系的Pn值较高而冀5418CHA不育系Pn值较低；（4）不同不育类型的光合产物积累存在～定差异，K、V、T型3种不育系的千粒重为29．1～37．82g，CHA型不育系仅22．73～23．51g，均低于保持系，在淀粉和可溶性糖含量上也有类似的表现，但是CHA型不育系蛋白质含量为19．13％-19．93％，高于其他类型不育系0．58％-3．08％，保持系最低，仅15．47％～15．38％，说明CHA造成的伤害对光合产物积累造成的影响要大于胞质效应；（5）不同基因型对细胞质的反应不同。冀5418不育系与其B系在光合速率及光合产物积累上的增减幅度小于太911289，说明后者对胞质反应较前者敏感。

K、T、V、CHA型杂种小麦品质性状的细胞质效应

采用 2个母本基因型、4种细胞质类型的两因素实验设计 ,对T、K、V、CHA型杂种小麦的品质性状表现及其差异进行了比较研究。结果表明 ,K、V、T型细胞质对杂种小麦某些品质性状具有正向效应 ,其中K型细胞质效应较大 ,其杂种的千粒重、面粉白度、蛋白质含量、湿面筋含量、面筋指数、沉淀值、降落值、面团形成时间、稳定时间和评价值均表现正向杂种优势 ,超MP 2 .0 8%～ 16 .82 % ,与V、T、CHA型相比有多个性状优势的差异达到显著水平 ;容重和吸水率的杂种优势在细胞质间差异不显著 ,无细胞质优势效应 ;母本基因型与细胞质具有交互作用 ,对杂种F1 代的性状表现具有一定影响。杂种F1 的容重和评价值的互作值较大 ,分别为 2～ 34g·L- 1 和 2～ 2 8,蛋白质含量和稳定时间的互作值较小 ,分别为0 .4 2 %～ 2 .4 %和 0 .2～ 2 .2min。对优势效应大的细胞质类型 ,选择敏感的母本基因型配制杂种有利于提高品质。

棉花晋A细胞质雄性不育系酶活性的研究

酶活性的变化与育性有着密切的关系。对与线粒体有关的过氧化物酶、超氧化物歧化酶、细胞色素氧化酶和琥珀酸脱氢酶进行了酶活性分析。结果表明:在花粉发育的各个时期晋A不育系的酶活性比其保持系明显要低。这表明了不育系与其保持系之间体内代谢强度的差异,也说明酶活性的变化与雄性不育现象有着直接的关系。

大豆质核互作雄性不育系W_(945)A、W_(948)A的选育

用大豆Ｗ２０２（中油８９Ｂ）作母本，分别与Ｗ２１０、Ｗ２１２二个品种杂交的Ｆ１ 发现不育株，通过正反交测定属质核互作型不育，通过连续四代核置换回交，获雌雄育性较稳定的二个不育系Ｗ９４５Ａ和Ｗ９４８Ａ。

甘蓝型油菜胞质雄性不育系C MS22A的选育与遗传 I.双低不育系C MS22A的选育及其败育类型研究

甘蓝型双低油菜CMS2 2A是河南省农业科学院棉花油料作物研究所利用 384A中的雄性不育源、采用单株成对连续回交的方法育成的双低油菜细胞质雄性不育系。其突出表现为雄性不育彻底且遗传稳定 ,配合力高 ,抗病抗倒 ,品质优良。CMS2 2A属典型的败药型雄性不育系 ,雄蕊败育短缩至雌蕊基部 ,♂ /♀ <0 .5 ,花药退化变小、无花粉。雌蕊发育正常 ,花瓣虽变小皱缩 ,但花瓣开张正常 ,可以正常接受外来花粉、受精结实。不育率稳定保持在 99%以上。

高粱A1型细胞质雄性不育系与保持系线粒体基因组分析比较

线粒体基因组易位是导致作物细胞质雄性不育(Cytoplasmic male sterility,CMS)性状产生的重要遗传机制。比较高粱A1型细胞质雄性不育系与保持系线粒体基因组,寻找易位区为克隆高粱A1型细胞质雄性不育相关基因奠定基础。以高粱A1型细胞质雄性不育系Tx623A和其保持系Tx623B为试验材料,采用二代Illumina Hiseq结合三代PacBio测序技术,对2个样品的线粒体基因组进行组装,比较和分析不育系和保持系基因组结构和基因差异。高粱Tx623A和Tx623B线粒体基因组大小分别为449 727 bp和452 772 bp,预测编码开放阅读框(Open reading frame,ORFs)分别为147和145个,且两基因组特有基因分别为8个和6个。两线粒体基因组共线性比较分析,发现存在一个57 kb的基因组片段易位的结构变异(Structural variation,SV)区域,该易位区可能与A1型细胞质雄性不育有关。Tx623A和Tx623B线粒体基因组中易位区为高粱A1型细胞质雄性不育基因克隆提供了基因组信息。

甘蓝型油菜NEA胞质不育双低恢复系选育研究

NEA是笔者所在课题组在远缘杂交的辐射诱变后代中发现的一类甘蓝型油菜胞质不育新材料,叶色深绿,营养体旺,不育性稳定彻底,群体不育株率和单株不育度均达100%,完全不同于核不育、Pol-cms和陕2A。恢复源的寻找是NEA育种利用的关键。利用丰富的油菜地方种质资源、创新资源及引进资源,采取辐射诱变和广泛测交方式,发现了能恢复NEA育性的基础材料,进一步的遗传研究表明,恢复NEA育性的是细胞核内一对显性基因。利用含恢复基因的基础材料,进行广泛转育,正反杂交、连续回交、复合杂交、聚敛杂交、测交、连续自交、配合力测定、单株品质测试,选育出一系列稳定纯合的NEA胞质双低恢复系,如NR67、NR722、NR403、NR1917、NR9482。

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。

普通小麦细胞质雄性不育系及其保持系线粒体DNA的RAPD分析

应用随机扩增多态ＤＮＡ（ＲａｎｄｏｍａｍｐｌｉｆｉｅｄｐｏｌｙｍｏｒｐｈｉｃＤＮＡｓ，ＲＡＰＤ）技术，对普通小麦细胞质雄性不育系（Ａ）及其保持系（Ｂ）线粒体基因组（ｍｉｔｏｃｈｏｎｄｒｉａｌｇｅｎｏｍｅ）的指纹图谱进行了分析。共使用引物４１个，其中２０个引物得到了扩增产物，部分引物扩增结果表现多态性。说明线粒体ＤＮＡ（ｍｉｔｏｃｈｏｎｄｒｉａｌＤＮＡ，ｍｔＤＮＡ）序列在不育系和保持系之间存在差异，进一步明确了小麦细胞质雄性不育（ｃｙｔｏｐｌａｓｍｉｃｍａｌｅｓｔｅｒｉｌｅ，ＣＭＳ）与ｍｔＤＮＡ的关系。