甘蓝型油菜生态型细胞质雄性不育两用系的研究Ⅲ. 8-8112AB的温度敏感性及其遗传

8-8112AB是由甘蓝型油菜pol细胞质生态型雄性不育两用系AB1与双低甘蓝型油菜品种杂交选育出的新型双低生态型pol CMS两用系,其在武昌秋播时表现雄性可育,在兰州或秭归高山夏播时表现雄性不育.在相对低温条件下,8-8112AB与正常油菜品系1141B(pol CMS保持系)的花器官形态特征表现相似,而在相对高温条件下,则与pol CMS不育系1141     

油菜胞质不育类型相关基因研究进展

细胞质雄性不育在油菜杂种优势利用中具有重要作用,研究细胞质雄性不育的不育机理对有效利用杂种优势,实现三系配套及创造强优势组合具有重要意义。介绍Ogu CMS,Pol CMS,Nap CMS等目前国际上主要的甘蓝型油菜细胞质雄性不育类型,综述了线粒体基因组上与CMS性状相关基因的位点或片段的最新研究进展,并讨论当今学术界有关CMS产生机理的主要论点。     

甘蓝型油菜胞质雄性不育系22A的遗传研究

甘蓝型双低油菜CMS22A是河南省农科院棉油所利用384A中的雄性不育源、采用单株成对连续回交的方法育成的新型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。对CMS22A育性遗传规律研究表明,该不育系属典型的败药型雄性不育系,其雄性不育的遗传受不育细胞质(S)和一对隐性核不育主效基因(rr)的共同控制,属孢子体不育,基因型为S(rr)。     

甘蓝型油菜萝卜细胞质雄性不育杂种优势利用研究进展

甘蓝型油菜萝卜细胞质雄性不育系统的不育系育性稳定、彻底,目前被认为是油菜杂种优势利用最为安全、有效的细胞质雄性不育类型。本研究主要针对甘蓝型油菜萝卜质不育系统的Ogura CMS和Kosena CMS两种主要类型,从其转育与改良、育性相关位点及其恢复基因的定位和克隆三个方面,综述了近年来关于萝卜质不育系统的相关分子机理及在甘蓝型油菜杂种优势利用上的主要研究进展。本研究最后还探讨了该研究领域目前存在的问题,并提出了中国甘蓝型油菜萝卜细胞质雄性不育系统研究的发展方向。     

春油菜区 Pol CMS 油菜杂交制种技术

Pol CMS是指波里马细胞质雄性不育,是油菜杂种优势利用的主要途径。从基地选择、播种期确定、播种方法、水肥管理、病虫草害防治、花期调节、除杂去劣、辅助授粉,收获贮存等方面,总结了Pol CMS油菜品种在青海、甘肃等春油菜产区实施杂交制种的技术要点,为波里马细胞质雄性不育杂种优势利用系统杂交油菜种子高质高产生产提供了技术参考。     

植物细胞质雄性不育基因的鉴定及育性调控机理

细胞质雄性不育（cytoplasmic male sterility,CMS）是作物杂交种生产的主要授粉控制系统,研究细胞质雄性不育分子基础及调控机理对利用杂种优势提高作物产量具有重要的指导意义。本文从植物的线粒体基因与细胞质雄性不育的关系着手,列举了植物CMS基因的鉴定情况,重点介绍了研究较多的矮牵牛、玉米、水稻和油菜细胞质雄性不育基因的鉴定进展及其对不育性状的调控。同时根据恢复基因与不育基因的相互作用情况阐述了育性恢复的可能机理,并对植物CMS分子机理的研究前景进行了展望。     

一个甘蓝型油菜细胞质雄性不育相关基因克隆和表达分析

利用甘蓝型油菜与播娘蒿融合杂种中的不育株与甘蓝型油菜杂交、回交,得到细胞质雄性不育系NJ65A。根据pol和nap雄性不育高度相关的基因orf224和orf222设计兼并引物,从NJ65A植株中克隆得到一个长度为675bp的基因,命名为orf224-NJ65A。该基因编码224个氨基酸,氨基酸序列与orf224、orf222和orf220序列相似性分别为：70％、53％、60％。RT—PCR表达分析表明：orf224-NJ65A在所检测的组织中都有表达,但在花、花蕾和茎中的表达量较大。推测orf224-NJ65A与细胞质雄性不育有关。     

高粱A2型细胞质雄性不育系小孢子发生的细胞学观察和减数分裂染色体行为分析

高粱凡型细胞质雄性不育性（CMS）的细胞质来源于IS12662C，A2细胞质杂交种目前已用于生产。本文以A2／B2 V4为材料，对A2CMS小孢子败育过程作了细胞学观察，并对小孢子败育过程中减数分裂的染色体行为作了分析。研究发现，在A2雄性不育系A2V4的花药发育过程中，绒毡层细胞不形成或提前解体；绒毡层细胞畸形化；绒毡层细胞虽发育正常，但小孢子母细胞减数分裂行为异常；这些都导致小孢子退化。A2细胞质雄性不育花粉母细胞减数分裂行为从后期Ⅰ开始出现异常，同源或姊妹染色体向两极分离时滞后或不分裂；染色体多倍化；一个细胞内出现多核和多核仁现象，最终导致小孢子败育。     

甘蓝型油菜细胞质雄性不育系Polima A和陕2A的研究进展

Polima A和陕2A是两个经典实用的甘蓝型油菜细胞质雄性不育(cytoplasmic male sterility,CMS)系,这两个著名的不育系在中国乃至世界油菜杂种优势的研究和利用中发挥了重要作用.有关这两个不育系的研究,过去主要集中在形态解剖学、细胞学、生理生化、遗传学以及杂种优势的利用等方面,近年来,主要集中在分子生物学方面.本文从以上方面对PolimaA和陕2A的国内外研究状况进行了综述,介绍了近年来这两个不育系的最新研究进展,以及两个不育系在生产实践中的应用、存在的问题和解决的途径.随着分子生物学技术的不断发展,对这两个不育系的研究将会更加广泛和深入.有关Polima A和陕2A的深入研究,对进一步探索油菜CMS的分子机理,更好地利用这两个不育系为我国油菜科研和生产服务有重要意义.     

提高甘蓝型油菜细胞质雄性不育杂交制种纯度的技术研究

用细胞质雄性不育系(CMS)陕2 A的微粉授予不育系陕2 A的柱头,在4 h和6 h后,再分别授以恢复系垦C1的花粉,其异交率仍可达73%和53.7%,据此作者提出了在花粉授精过程中同品种花粉的迟钝性,异品种花粉的优先性理论;同时还提出提高父母本授粉位差、缩小父母本行距、增加父本花粉量、加强辅助授粉、重复授粉等技术措施,可有效控制细胞质雄性不育微粉给油菜杂交制种造成的不良影响,对提高制种纯度有重要参考价值.     

Breeding a genie and cytoplasmic double-MS line of rapeseed (Brassica napus L.)

All rapeseed lines with Polima male sterility (MS) that are applied in hybrid cultivars have the problem that their sterility varies with temperature. To overcome this problem, two double-MS lines with genie (GMS) as well as cytoplasmic male sterility (CMS) genes were synthesized through seven generations of breeding, based on a systematic study of changes in fertility of the genie and cytoplasmic male sterility lines. The fertility of the new sterility lines was determined by observation of the floral organs and by pollen staining. The results showed that, in the double-MS lines, half the plants maintained the features of the Polima CMS line, while the other half behaved like the GMS line. The GMS genes were correctly expressed in the Pol cytoplasm, but there was little interaction between the GMS and CMS genetic systems.     

油菜野芥NSa细胞质雄性不育系的特异性分子鉴定

常规细胞质类型鉴定方法花费时间长、工作量大、鉴定效率低、应用性较差。基于基因组重复序列的PCR技术(rep-PCR)在重复序列丰富的线粒体和原核生物的基因组鉴定中效果较好, 本研究对8份油菜资源的细胞质进行分子鉴定,结果表明rep-PCR可以明确区分甘蓝型油菜中常见的6种雄性不育细胞质。同时对甘蓝型油菜和野芥(Sinapis arvensis)体细胞杂交创建的、育性较为稳定的NSa野芥雄性不育胞质进行了分子特异性鉴定,获得了两条NSa不育胞质的特征DNA条带。但波里马和萝卜质不育细胞质的特异性引物在NSa中不能扩增出特异性片段,说明NSa不属于这两种细胞质类型。本研究结果为该不育胞质系统的利用和知识产权保护提供了理论依据和技术支撑。     

双低甘蓝型油菜杂交种亲本指纹图谱构建和杂交种纯度鉴定

利用SSR分子标记构建了18份恢复系和21份不育系共39份甘蓝型油菜杂交种亲本的DNA指纹图谱,筛选出H9901和H9905两个杂交种亲本的共显性SSR标记Na10G08,并用其鉴定了H9901、H9905两个大田生产F1杂种群体的纯度。对比SSR和田间调查的鉴定结果表明,SSR标记鉴定可以识别更多的非杂交种单株,如母本因微粉导致的自交、花粉污染、     

Cytoplasmic male sterility with self-incompatibility,a novel approach to utilizing heterosis in rapeseed (<Emphasis Type="Italic">Brassica napus</Emphasis> L.)

The Polima cytoplasmic male sterility (CMS) system has been successfully used in three/two-line hybrid production in rapeseed (Brassica napus L.). However, the sterility of the Polima (pol) CMS lines is sensitive to temperature fluctuations. Also, traces of pollen can cause self-pollination within the CMS lines, which results in reduced levels of F₁ hybrid seed purity and leads to a significant yield loss. Self-incompatibility (SI) is another important approach for hybrid seed production in rapeseed. Despite having a wide range of restorers and being easily selected in a breeding program, SI system has some drawbacks. In this study, SI genes from a self-incompatible line of Brassica napus were transferred to a pol CMS line and S372A, a novel line of combined cytoplasmic male sterility with self-incompatibility was bred. Due to the SI genes, this line produced very few seeds when it was selfed at low temperature and no seeds at high temperature. This suggested that the line with CMS + SI had combined the advantages and overcome the disadvantages of both the pol CMS and SI systems. Furthermore, our results showed that most of the maintainers and all the restorers of the pol CMS system were also maintainers and restorers of the CMS + SI line, respectively. This indicates that the CMS + SI system can be easily used to establish three-line hybrids of rapeseed, and we believe this novel system could be extended to other species of Brassica.     

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。     

Cytological and molecular characterization of a new cytoplasmic male sterility in rapeseed

It is very important for rapeseed hybrid production to develop and utilize a novel cytoplasmic male sterility (CMS) system concerning the possible risk because of a narrow cytoplasm background. Here the anatomy of anther development in the CMS system, named NCa , was observed using light microscope and transmission electron microscope, and the restriction fragment length polymorphism (RFLP) analyses of mitochondrial DNA of NCa sterile line were also performed in comparison with the other rapeseed sterile lines, such as pol , nap , ogura , and tour . The anther abortion of this CMS line occurred at the later uninucleate microspore stage, and the anatomic aborting characteristics were obviously different from all the other rapeseed CMS lines reported before. The RFLP analyses revealed that five probe/enzyme combinations could distinguish the five CMS lines. The results of anatomic observations and mitochondrial DNA polymorphism indicated that the NCa CMS system is a novel one which differs from the pol , nap , ogura , and tour systems.     

甘蓝型双低油菜胞质雄性不育系384A不育性的表现与遗传

对甘蓝型双低油菜细胞质雄性不育系３８４Ａ的不育性表现、遗传方式等方面的研究结果表明,该不育系属败药型不育,其败育彻底,育性稳定,花瓣开张正常,不育性的遗传受不育细胞质（Ｓ）和一对隐性核不育主效基因（ｒｒ）共同控制,属孢子体不育,基因型为Ｓ（ｒｒ）,同时也受若干位点上微效基因的影响。     

Establishment of a random-mating population of 'Polima' CMS restorers in Brassica napus by use of dominant genie male sterility

Five restorers of ‘polima’ cytoplasmic male sterility (pol CMS) cannot restore the fertility in dominant genie male sterility (DGMS). A dominant male sterility gene from both, a DGMS line Rs l046AB and DGMS hybrid ‘Zhongza No. 3’, was successfully introduced into Polima cytoplasm. A random-mating population of pol CMS restorers was established by using many double-low pol CMS restorers as pollinators to cross continuously to the DGMS plants which had Polima cytoplasm.     

Cytological observation of anther structure and genetic investigation of a new type of cytoplasmic male sterile 0A193‐CMS in Brassica rapa L.

Cytoplasmic male sterility (CMS), a maternally transmitted failure in pollen formation, is an effective pollination control system in hybrid rapeseed (Brassica napus) breeding. However, CMS is not widely used in the related oilseed species Brassica rapa. In the past years, several male sterile plants have been isolated from the B. rapa landrace ‘0A193’, collected in Shaanxi, China, in 2011. It is noteworthy that the fertility expression of 0A193‐CMS was affected by temperature. In contrast to pol CMS, fertility tests with 18 B. rapa and 9 B. napus accessions suggest that a different system of maintaining and restoring is responsible for the observed phenotype. Further on, genetic investigation evidenced that fertility of 0A193‐CMS is controlled by both cytoplasmic and one pair of nuclear recessive genes. Interestingly, plants of the 0A193‐CMS type possess a highly specific fragment of the mitochondrial gene orf222, a crucial regulator of male sterility in nap CMS. Our study broadens the CMS resources in B. rapa and provides a highly applicable alternative to pol CMS and ogu CMS for hybrid breeding production.