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。     

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。     

作物雄性不育细胞质遗传效应的研究

细胞质雄性不育,即质核互作雄性不育是细胞质与细胞核相互作用产生的,是作物三系法杂种优势利用的基础。自本世纪初,随着多种作物细胞质雄性不育的发现以及杂种优势利用研究的开展,人们对于雄性不育细胞质进行了广泛的研究。近年来,我国科技工作者在这个方面的工作也取得了很大进展。本文主要综述水稻和小麦雄性不育细胞质遗传效应的研究。     

体细胞杂交在油菜细胞质雄性不育创建和改良中的应用

油菜细胞质雄性不育作为生产油菜杂交种的主要授粉控制系统得到广泛利用，但由于油菜种内自然发生的有利用价值的雄性不育细胞质类型不多，所以种属间转移现有不育细胞质及发掘新型不育细胞质意义重大。体细胞杂交不仅避开了有性杂交亲和障碍的限制，还可实现两个杂交亲本细胞质基因组的重组，是一种快速有效、应用广泛的转移和诱导雄性不育细胞质的手段。本文综述了利用原生质体杂交技术在油菜种内、种间和属间转移雄性不育细胞质以及通过细胞质基因组重组改良和创建新型雄性不育细胞质的研究进展，并讨论了利用体细胞杂交方法创建新型雄性不育细胞质的应用前景。     

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。     

秦油二号制种的防杂保纯

秦油2号是利用甘蓝型油菜细胞质雄性不育“三系”组配的杂交种,属典型的虫媒花,在制种中极易受昆虫和风力影响传粉,与其它油菜和十字花科作物串花引起生物学混杂,所以防杂保纯技术十分复杂。主要应围绕亲本、环境、媒介三个系统做好工作。     

油菜雄性不育分子机理研究进展

该文从分子生物学角度概括介绍了油菜雄性不育机理的最新研究进展，包括细胞质雄性不育与线粒体分子生物学；细胞质雄性不育与基因表达调控，细胞质雄性不育与核质基因在分子水平上的互作关系；细胞核雄性不育的分子机制，利用生物技术获得雄性不育系等。     

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。     

Ogura不育胞质对甘蓝型油菜与萝卜种间可交配性的影响

用三个甘蓝型油菜萝卜细胞质雄性不育系（Ogu CMS)及其相应的保持系作母本与萝卜杂交，以了解Ogura不育胞质对甘蓝型油菜与萝卜种间可交配性的影响。结果表明，Ogura不育细胞质对甘蓝型油菜与萝卜种间可交配性有正效应，可交配性受核-质互作控制。通过胚珠培养，获得了甘蓝型油菜萝卜细胞质雄性不育系与萝卜的属间杂种。     

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。     

人工合成具有白菜或甘蓝细胞质的甘蓝型油菜

为比较甘蓝型油菜不同细胞质基因组的遗传效应及其与核基因的相互作用,对12个白菜(2n=20)与1个芥蓝(2n=18)的正反交种间杂种分别进行子房和胚培养,人工合成白菜和甘蓝细胞质甘蓝型油菜。结果表明,白菜×芥蓝的正交子房培养杂种苗平均诱导率2.32%,甘蓝×白菜的反交胚培养杂种苗平均诱导率为1.16%。不同杂交组合之间,杂种苗诱导率差异大,但相同亲本正反交杂种获得的难易趋势相似。将单倍体杂种小苗在含0.01%秋水仙碱的MS培养基中预培养处理10d的染色体加倍效果最好,加倍率达59.32%。人工合成的甘蓝型油菜农艺性状类似于栽培甘蓝型油菜,无论正交或反交合成的甘蓝型油菜,其农艺性状介于父母本之间,但更接近于母本。合成油菜花粉育性在40.53%～88.95%之间。     

A cytogenetic study of the progenies of hybrids between Brassica napus and Brassica oleracea, Brassica bourgeaui, Brassica cretica and Brassica montana

In this cytogenetic study the progeny of all crosses were investigated in F₁, F₂ and backcross (BC₁) hybrids. Brassica napus and F₁ hybrids between B. napus and B. oleracea, and between B. napus and three wild relatives of B. oleracea (B. bourgeaui, B. cretica and B. montana). Each of the wild relatives has 18 somatic chromosomes. Interspecific F₁ hybrids were obtained through ovary culture mean. These had 28 and 37 chromosomes and their mean pollen fertility was 10.7% and 93.0%, respectively. Many F₂ and BC₁ seeds were harvested from the F₁ hybrids with 37 chromosomes after self‐pollination and open pollination of the F₁ hybrids, and backcrossing with B. napus. Many aneuploids were obtained in the F₂ and BC₁ plants. It is evident from these investigations that the F₁ hybrids may serve as bridge plants to improve B. napus and other Brassica crops.     

Interspecific hybridization between Brassica carinata and Brassica rapa

The crossability between Brassica carinata (BBCC, 2n=34) and Brassica rapa (AA, 2n=20), and the cytomorphology of their F₁ hybrids were studied. Hybrids between these two species were only obtained when B. carinata was used as the female parent. The hybrid plants exhibited intermediate leaf and flower morphology, and were found to be free from white rust and Alternaria blight diseases. One of the four F₁ plants was completely male sterile, while the remaining plants had 4.8, 8.6, and 10.9% stainable pollen, respectively. No seed was produced on hybrid plants under self pollination or in backcrosses; but seed was obtained from open pollination. The occurrence of the maximum of 11 bivalents as well as up to 44.8%) of cells with multivalent associations in the form of trivalents (0‐2) and a quadrivalent (0‐1) in the trigenomic triploid hybrid (ABC, 2n = 27) revealed intergenomic homoeology among the A, B and C genomes. Meiotic analysis of F₁ hybrids indicated that traits of economic importance, such as disease resistance, could be transferred from B. carinata to B. rapa through interspecific crosses.     

NaCl胁迫对甘蓝、白菜和油菜种子萌发的影响

为研究蔬菜种子吸收或外排Na+能力与其耐盐性的关系,采用非损伤微测技术检测了628甘蓝、青杂中丰白菜、苏州青油菜、茼蒿、上海鸡毛菜种子外Na+流速的动态变化.结果表明,在10mmol/L NaCl溶液处理下,甘蓝和茼蒿种子外排Na+,而白菜、油菜和鸡毛菜种子吸收Na+,其中,甘蓝、白菜和油菜种子对Na+吸收或外排的效果明显.随后以甘蓝、白菜和油菜种子为材料,在不同NaCl浓度下对种子萌发进行了实验,结果表明,随着NaCl浓度升高,甘蓝、白菜和油菜种子的发芽率、相对发芽率、发芽势和发芽指数都呈明显下降趋势.当NaCl浓度达到300mmol/L时,甘蓝、白菜和油菜种子的发芽率分别为37.33％、14.67％和10.00％,耐盐半致死浓度分别为323.73,241.68mmol/L和200mmol/L.初步说明种子可以通过增加Na+的外排,减弱高浓度Na+对种子的毒害作用.     

Hybridization in Brassica juncea × Brassica campestris through ovary culture

Summary Ovary culture has been employed for the production of interspecific hybrids of a partially compatible cross of Brassica juncea (2n=36) × Brassica campestris (2n=20). Five to seven days old ovaries cultured on White's medium supplemented with casein hydrolysate (300 mg/l) and sucrose (5%) produced more seeds than any other media tried, but seed development was better on media fortified with plant hormones. The seed yield was better in B. juncea × B. campestris than their reciprocal cross. The plants obtained from ovary-derived seeds were transferred to the field; they were intermediate in some morphological characters and chromosome number (2n=28) as compared to their parents. The flower buds generally did not open and had poorly developed anthers with mostly sterile pollen. The pod size/setting was very much reduced, but healthy seeds were obtained.     

Transgene introgression from Brassica napus to different varieties of Brassica juncea

Transgene introgression from transgenic rapeseed (Brassica napus) to different varieties of B. juncea was assessed in this study. Crossability between a transgenic rapeseed line Z7B10 (pollen donor) and 80 cultivars of 16 B. juncea varieties (including two wild accessions) was estimated by artificial pollination in a greenhouse. As a result, interspecific crossability between the transgenic Z7B10 line and the 80 B. juncea cultivars varied considerably, with seeds per flower from 0.00–10.67. Seed germination rates of the interspecific F₁ hybrids ranged from 49.0%–89.3%. The estimated frequencies of natural gene flow from the transgenic Z7B10 line to 10 B. juncea cultivars with different uses in the experiment field varied from 0.08% to 0.93%. The natural F₁ hybrids were highly sterile, with seeds per silique ranging from 0.27 to 1.03. In addition, seeds per flower of hybrid descendants varied from 0.02 to 0.22 when F₁ hybrids were self‐pollinated, and those ranged from 0.03 to 0.30 when F₁ hybrids were backcrossed with their corresponding B. juncea parents. Results of this study suggest a low level of transgene introgression from transgenic rapeseed to different B. juncea varieties, which provides a sound scientific basis for the safety management of coexisting transgenic B. napus and B. juncea varieties in China.     

以NIRS定量分析甘蓝型油菜、芥菜型油菜饼粕赖氨酸含量

作物品质育种工作需要快速和准确,并且不破坏种子的情况下来测定有关参数,但传统的品质鉴定方法在分析速度和效率以及种子的留存上存在局限性,难以满足上述要求.近红外反射光谱分析技术(near infrared reflectance spectroscopy,NIRS)是一种不需要对种子进行前处理,即可得到几乎全部农产品品质参数的分析技术[1].NIRS技术现已广泛应用于农产品[2]、食品[3]、果汁[4]、饲料[5]等的分析检测.我国该技术的应用虽起步较晚,但在农作物品质分析上已取得了一些可喜的进展[6,7].用近红外仪测试油菜种子的含油量、饼粕的蛋白质含量与硫甙含量,已基本接近和达到化学分析所需求的精度[8].     

Interspecific transfer of Brassica juncea-type high blackleg resistance to Brassica napus

Summary Complete resistance to Leptosphaeria maculans, the cause of blackleg of oilseed rape (Brassica napus), was transferred from B. juncea to B. napus through an interspecific cross. B. juncea-type complete resistance (JR) was recognized first in one F₃ progeny (Onap^JR) by the absence of leaf-lesions on seedlings and canker-free adult plants. The commercially important characters of B. napus were retained in advanced lines of Onap^JR, which combined JR with low erucic acid levels (<0.5%), high seed yield and variable maturity dates.JR appeared to be inherited as a major gene or genes. Segregation for resistance and susceptibility contintied to occur during later generations of selection of Onap^JR. JR was readily transferred from Onap^JR to other suitable B. napus cultivars or lines with partial resistance to blackleg and resulted in highly vigorous carly generation selections adapted to cold, wet situations along with complete resistance to blackleg.     

甘蓝、大白菜和甘蓝型油菜EXO70A1基因的克隆与表达特性

从甘蓝、大白菜与甘蓝型油菜中分离出EXO70A1基因,对该基因的序列进行生物信息学分析, 然后转化酵母Y187, 应用半定量RT-PCR检测BoEXO70A1基因的表达特性。结果表明, 3种芸薹属植物EXO70A1序列长度均为1 917 bp,相似性97.1%, 它们的gDNA序列均为单一序列,长度分别为3 797、3 752和3 770 bp,一致度达91.0%,均由12个外显子及11个内含子组成,除了第4、第5、第6、第8个内含子外,其余内含子的保守性低于外显子; 推导的3种蛋白质序列(BnEXO70A1、BrEXO70A1和BoEXO70A1)的相似度与一致性分别达99.8%与98.1%,其二级结构、三维结构及理化特性高度相似。EXO70A1基因的11个内含子的剪切位点均符合“GU-AG”法则,剪切受体(AG)的前20~50个碱基存在一段保守的序列“CU(A/G)A(C/U)”; 3种芸薹属植物与拟南芥EXO70A1基因的12个外显子的对应序列长度完全相同,所构成的编码区的序列一致性达90.1%,相应的蛋白质序列的相似度与一致性分别达99.8%与93.7%; 分子进化分析表明, EXO70A1在整个EXO70蛋白家族中及不同的植物间表现出较高的保守性; BoEXO70A1在酵母细胞Y187呈现弱表达; EXO70A1在甘蓝的雄蕊、幼茎、幼嫩花瓣、雌蕊、幼根及叶片中均能表达,可能属于组成型表达基因,但是其表达量在不同发育时期的不同器官中存在差异,授粉前雌蕊中最高,雄蕊中最低。由此可知,EXO70A1在芸薹属植物中整体高度保守, 但在酵母转化株和甘蓝各器官中的组成型表达有所差异,推测EXO70A1在植物细胞中具有多种重要的功能。