Identification of a Stable Maintainer Line for 'Polima' Cytoplasmic Male Sterility in Rapeseed (B. napus L.)

‘Polima’ cytoplasmic male sterility (CMS) was transferred to three different genotypes of B. napus i.e. GSL-1, ISN706 and HNS-8 by repeated backcrossing and was found to be completely stable in the nuclear background of synthetic B. napus genotype, ISN706, derived from a cross B. campestris ssp. oleifera var. brown sarson ×B. oleracea var. botrytis cv. Tusa Katki'. The BC₅ and BC₆ generation lines of ISN706, GSL-1 and HNS-8 with ‘Polima’ CMS were grown under a range of temperature and photo-period conditions to test their stability. No breakdown in sterility was observed in ‘Polima’ ISN706 and, therefore, this genotype can be used as a maintainer for hybrid seed production.     

Identification and Inheritance of Fertility Restorer Genes for 'tour' CMS in Rapeseed (Brassica napus L.)

Eighteen genotypes of Brassica napus were crossed to a cytoplasmic male sterile (CMS) line of B. napus BO 15 carrying B. tournefortii cytoplasm (‘tour’ cytoplasm). Fourteen genotypes were found to be stable maintainers of the ‘tour’ CMS. Of the remaining four genotypes, GSL-1 and ‘Asahi-natane’ were found to be heterozygous and ‘Mangun’ and ‘Yudal’ were homozygous for the restorer gene. Analysis of the F₁ and F₂ progenies of (CMS) BO 15 בMangun’ and (CMS) BO 15 בYudal’ showed that fertility restoration is controlled by a single dominant gene. The availability of a number of stable maintainer lines and the simple inheritance pattern of fertility restorer gene makes ‘tour’ CMS a useful system for hybrid seed production in rapeseed.     

The Inheritance of Polima Cytoplasmic Male Sterility in Brassica napus L.

The restores tested in this study have only one pair of fertility-restoring genes (Rf) but many temperature-sensitive genes (Ts). The Rf genes are alleles at the same locus. The cytoplasm of the restorers RC₁, RC₂, RC₃ and ‘Huaie’ is male sterile, but that of RC₄, is normal.     

特早熟甘蓝型春油菜恢复系核心种质构建

通过对特早熟甘蓝型春油菜恢复系种质资源进行评价和利用,降低育种工作中恢复系与不育系选配杂交组合的工作量,从而更好地为杂交种选育服务。通过对97份波里马雄性不育系的恢复系进行简化基因组（SLAF-seq）测序,开发SNP标记,进而分析群体遗传关系,构建核心种质群体。结果显示,共获得527 872个SLAF标签,842 248个有效SNP;利用有效SNP对97份种质资源进行聚类分析,可将其分为5类;利用Core Hunter构建核心种质,构建了C30核心种质29份,C40核心种质38份,C30和C40核心种质平均等位基因覆盖度分别为99.89%和99.96%,这些材料能够代表整个资源的变异程度,达到了构建核心种质的要求。     

Studies on "Three Line" Polima Cytoplasmic Male Sterility Developed in Brassica napus L.

Polima CMS lines can be divided into three groups according to the sensitivity of male sterility to temperature: 1) high temperature CMS lines (62.9%), 2) low temperature CMS lines (28.6 %) and 3) stable CMS lines (8.5 %). The heterosis of Pol CMS hybrids is evident and common in some of the characters tested, especially in plant seed yield and siliqua number. The cytoplasm has no deleterious effects on the FI characters tested.     

Tournefortii male sterility system in Brassica napus. Identification, expression and genetic characterization of male fertility restorers

A germplasm collection of 152 diverse rapeseed accessions from Canada, China, France, India, Poland and South Korea was assayed for identifying new fertility restorers and sterility maintainers for a Tournefortii (tour) cytoplasmic male sterility (CMS) system in rape‐seed. Only 16 (10.5%) genotypes showed complete fertility restoration following hybridization with tour CMS line NE 409A. Notable among these were GSL 8851, GSL 8953, Mokpo # 9, Mali, Buk‐wuk‐13, Kuju‐27 and Mokpo # 84. As many as 78 (51.3%) genotypes were perfect maintainers of sterility, the remaining 58 (38.2%) genotypes were classified as partial maintainers. To study the inheritance of fertility restoration, 20 CMS (tour) rapeseed lines were crossed with the four best fertility restorers, namely GSL 8851, GSL 8953, Kuju‐27 and Mokpo # 9, to obtain F₂ and test cross populations. Segregation data indicated that fertility restoration for tour CMS was governed by two genes, of which, one is stronger than the other (χ²_12:3:1). Differences in gene interactions were also observed (χ²_9:3:4) which could be explained on the basis of influence of female parent genotypes/or modified expression of the restorer gene(s) in different genetic backgrounds. Tests of allelism indicated that the restorer genes present in the four restorers evaluated were allelic.     

甘蓝型油菜细胞质雄性不育恢复基因的遗传研究

通过多年大量的筛选 ,在不同来源甘蓝型油菜中发现了一批恢复系 ,并利用筛选的恢复系中的恢复基因 ,转育出一批恢复性能良好、品质优良、配合力高的恢复系。对这些恢复系的遗传研究结果表明 ,这些不同来源的恢复基因为一对主效基因 ,但这些不同来源的恢复基因的等位性不同     

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。     

中国甘蓝型油菜细胞质雄性不育的主要类型及在育种实践中的应用

我国甘蓝型油菜细胞质雄性不育的类型主要有：ogura CMS、nap CMS、pol CMS、Shan 2A CMS、Hau CMS、NCa CMS、Nsa CMS和NEA CMS。pol CMS和Shan 2A CMS在我国油菜杂种优势利用中发挥着重要的作用。新型不育胞质的育成以及在育种上的应用,不仅丰富了细胞质雄性不育种质资源,而且为我国油菜产业可持续发展提供了保证。     

Independent Inheritance of Flower Colour and Male-Fertility Restorer Characters in Brassica napus L.

Available material of oilseed (Brassica napus L., AACC) comprises two yellow-flowered breeding lines and a white/pale-flowered line of resynthesized rape. The flower colour white/pale is dominant over yellow, and is controlled by a gene located in the C-genome. The yellow-flowered genotypes acted as restorer lines and the white/pale-flowered genotype as a maintainer line in a cytoplasmic male sterility system. The segregation pattern of flower colour and male fertility restorer characters were studied in F₂ generations of crosses between these lines, also in a three-way cross additionally including a yellow flowered B. campestris (AA) line. Evidense was obtained in support of the conclusion that the flower colour and male fertility restorer characters are monogenically controlled and independently inherited. Whether the male fertility restorer gene is located in the A or C genome remains to be determined.     

甘蓝型油菜Pol CMS育性恢复基因的RAPD标记

采用恢、 保回交群体和集团分离分析(BSA), 筛选了860个10 mer随机引物, 找到了与甘蓝型油菜波里马细胞质雄性不育系(Pol cms)育性恢复基因(Rf)连锁的两个RAPD标记AH19690和AI16830。 它们位于Rf的一侧,与该基因的遗传图距分别为5.9 cM和13.6 cM,两标记间的遗传图距为7.8 cM。这两个RAPD标记的发掘,为进一步利用分子标记     

利用小孢子培养选育榨菜胞质甘蓝型油菜CMS恢复系研究

以榨菜胞质甘蓝型油菜恢复系材料“06 B033”为母本,优质油菜新品种(系)为父本配制7个杂交组合,并对其F1代进行小孢子培养.结果发现,不同父本的杂交组合在小孢子培养成胚率上表现极显著差异;对获得的DH群体进行对榨菜胞质甘蓝型油菜CMS的恢保性鉴定.结果表明,有5个DH株系对榨菜胞质甘蓝型油菜CMS表现良好的恢复性.     

甘蓝型油菜显性细胞核雄性不育基因的AFLP标记

用甘蓝型油菜双基因显性细胞核雄性不育系Rs1046A和欧洲油菜品种Samourai构建了一个回交分离群体。在群分法（BSA）构建的不育池和可育池中共筛选了256对AFLP引物组合,找到了与不育基因紧密连锁的两个AFLP标记(EA03MC1599和EA07MC01235), 它们与不育基因的遗传图距分别是3.5 cM和5.5 cM,而且位于不育基因的同一侧,标记间相     

油菜温敏雄性不育系373S的选育

发现和选育新型光温敏雄性不育材料,有利于深入揭示植物雄性不育的发生机制,丰富油菜杂种优势利用的方法。笔者报道一种新发现的环境敏感雄性不育系373S的选育过程及其育性随温度、日长的变化基本规律。2002年在一个天然雄性不育群体后代中发现1株嵌合不育株,经过连续自交,育成环境敏感不育系373S。2004—2006年在陕西杨凌进行分期播种试验,并进行了温度、光照脉冲处理试验。结果表明：373S是主要对温度敏感的雄性不育系,低温可育而高温不育,开花前2~9d的平均温度对育性影响显著,而日长对育性影响不明显,花蕾长度约2~3mm是育性转换的敏感时期。用一系列品种与373S测交,F1均为100%可育,在F2代温敏不育株出现几率很高,可以达到7.1%~14.6%。进一步精确分析373S光温反应特征、改良其农艺及品质性状、寻找适宜制种生态区的工作正在进行当中。     

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。     

甘蓝型油菜细胞质雄性不育材料RMcms的育性变化特性的研究

广恢型细胞质雄性不育材料(RMcms)在开花后期均有不同程度的微量花粉发生，主要分布于花序的上部，在同一植株上，其微粉发生的程度次序为主花序＞一次分枝＞二次分枝；微量花粉发生与开花前18天、19天或20天的平均日最高温和平均日均温有着密切的关系，当平均日最高温稳定通过20℃、平均日均温＞14．5℃时所开花朵即有不同程度微量花粉发生，说明该类型不育材料属高温敏感型。该类型不育系的不育性因细胞质的来源不同而有很大的差异，同一不育系群体中的不同植株的不育性也存在不同程度差别。     

Identification and Genetic Studies of the Inhibition of Dominant Male Sterility in Brassica napus

By transferring dominant male sterility (DMS), caused by the gene Ms, to genotypes with various types of cytoplasm 12 DMS lines were developed and a number of crosses made between the DMS lines and other genotypes of Brassica napus. During the course of this population improvement programme, 16 genotypes were identified as having the capacity to restore the fertility of F₁ plants with the Ms gene. According to pedigree analysis, the inhibitory gene in those lines probably originated from a few genotypes from Australia and Germany. In further studies the inheritance of the sterility inhibition was determined, providing definite evidence that dominant male sterility and its inhibition in B. napus are controlled by two dominant interacting genes rather than by multiple alleles.     

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.     

Inducing male sterility in Brassica napus L. by a sulphonylurea herbicide, tribenuron-methyl

A sulphonylurea herbicide, tribenuron‐methyl, methyl 2‐[[[[(4‐methoxy‐6‐methyl‐1,3,5‐triazin‐2‐yl) methylamino] carbonyl] amino] sulphonyl] benzoate, was used to induce male sterility in rapeseed. Application of 0.2 μg tribenuron‐methyl per plant at the bolting stage with the longest floral bud <2 mm and repeated 15 days afterwards, resulted in 94.5–100% plants being male sterile in six different breeding lines, but combined with low phytotoxicity. However, excessive double application of the chemical (>0.2 μg per plant) did have some significant impact on rapeseed, including stunting, fading leaves and petals, reduction in the size of floral parts and a shortened duration of flowering. The percentage of hybrid seeds from ‘84004‘ treated with 0.2 μg × 2 tribenuron‐methyl per plant and pollinated by a male parent ‘Huaye’ was 92.7%, which met the hybridity requirement in China. The results suggest that tribenuron‐methyl could be used as an efficient chemical hybridizing agent.     

Development of Polima temperature-sensitive cytoplasmic male sterile lines of Brassica napus through isolated microspore culture

A novel Pol CMS named Polima temperature‐sensitive cytoplasmic male sterile (Pol TCMS) is sterile at high temperatures and partially fertile at low temperatures. Its temperature sensitivity is controlled by several minor genes, and probably influenced by the genetic background, making it quite difficult to breed a stable Pol TCMS line through the conventional breeding methods. The present study aimed to obtain stable homozygous Pol TCMS lines by microspore culture. The highest frequency of embryogenesis was induced from cultures of buds sampled at 4–7 days before first blossom. The second method of microspore cultures mentioned in this study could significantly increase the production of embryos. Analysis of fatty acid composition, glucosinolate content and genetic diversity by simple sequence repeat (SSR) markers indicated that microspore culture was effective for selecting Pol TCMS lines.