结球甘蓝(Brassica oleracea var. capitata)迟抽薹基因SCAR标记转CAPS标记

为了更加精确结球甘蓝迟抽薹基因标记。本研究以结球甘蓝冬性强的迟抽薹材料‘P02’和冬性弱的易抽薹材料‘A21’杂交得F1代,F1代自交至F3代为试验材料,并以课题组所设计的特定序列扩增(sequence characterized amplified regions, SCAR)标记的SCN1/248为引物。在F3代的785株结球甘蓝中有711株扩增出目的条带,有74株未能扩增出目的条带。经测序,该序列长度为248 bp,运用该序列共设计3对酶切扩增多态性序列(cleaved amplified polymorphic sequence, CAPS)引物,其中1对引物均扩增出目的条带,多态性消失,另外两对分别命名为SC01和SC02。结合田间127株结球甘蓝抽薹性状调查,有65株表现为迟抽薹,59株表现为易抽薹,3株表现为中间型,与SCAR扩增检测结果基本一致,将SCAR标记转换成CAPS标记,建立CAPS标记体系。本研究进一步精化了分子标记辅助选择,为做精确定位功能基因,选育优良的迟抽薹结球甘蓝品种提供了理论依据。     

通过游离小孢子培养获得绿菜花（Brassica oleracea var.italica）再生植株

通过游离小孢子培养获得绿菜花（Ｂｒａｓｉｃａｏｌｅｒａｃｅａｖａｒ．ｉｔａｌｉｃａ）再生植株ＰｌａｎｔＲｅｇｅｎｅｒａｔｉｏｎｆｒｏｍＩｓｏｌａｔｅｄＭｉｃｒｏｓｐｏｒｅＣｕｌｔｕｒｅｏｆＢｒａｓｉｃａｏｌｅｒａｃｅａｖａｒ．ｉｔａｌｉｃａ７０年代初...     

Erucastrum abyssinicum × Brassica oleracea hybrids obtained by ovary and ovule culture

Pollen germination and pollen tube growth were studied in the cross Erucastrum abyssinicum × Brassica oleracea. 25 per cent ovules showed pollen tube entry into the micropyle 5 days after pollination. Embryos did not grow beyond the globular or heart-shaped stage. Thus post-fertilization barriers prevent hybridization in this cross. Sequential culture of ovaries and ovules 10 days after pollination helped rescue globular embryos. Three hybrids were obtained. All the hybrids flowered in vitro. Pollen fertility was 6.5 per cent. Leaf esterase and peroxidase isozymes confirmed the hybrid nature. Meiotic metaphase in the pollen mother cells showed 25 chromosomes. The presence of up to 9 bivalents and a quadrivalent indicates high homology between the two genera. The intergeneric hybrids have been multiplied using nodal segment culture. They are a source for obtaining cms cole crops and widening the gene pool of B. oleracea. This revised version was published online in July 2006 with corrections to the Cover Date.     

黑腐病对花椰菜（Brassica oleracea L. var. botrytis）幼苗根系形态和生理的影响

为了阐明黑腐病对苗期花椰菜的根系形态和生理的影响,对建立花椰菜抗黑腐病根系育种新途径提供依据,利用黑腐病菌感染花椰菜抗病品种雪峰的叶片后,研究植株根系发生的一系列形态和生理变化。结果表明,接种7天后的植株根长、根干重、地上部干重、根系活力比接种后2天的显著增加。接种处理的植株根长、根干重和地上部干重明显比未接种对照（CK）低,但是接种幼苗的根系活力比未接种对照的根系活力高。接种3天后,接种处理的根系可溶性蛋白质含量一直显著高于未接种对照;接种后0~4天,接种处理的根系超氧化物歧化酶（SOD）活性明显低于未接种对照,但到接种后5~7天,接种处理的根系SOD活性又高于未接种对照;根系多酚氧化酶（PPO）活性和脱落酸（ABA）含量在植株接种后都显著高于未接种对照。但是,黑腐病病原菌没有提高根系过氧化物酶（POD）活性。以上结果说明,黑腐病菌引起了花椰菜幼苗根部一系列形态变化,并通过诱导植株根系中防御酶活性和调控内源激素等抵御病原菌的进一步侵染。     

根癌农杆菌介导白细胞介素-4基因转化甘蓝(Brassica oleracea L.)研究

利用根癌农杆菌介导法,以白细胞介素-4(interleukin-4,IL-4)基因转化中甘11号甘蓝.本研究对可能影响il-4转化率的外植体类型、外植体的预培养时间、农杆菌的侵染时间以及外植体与农杆菌的共培养时间进行了优化.试验结果表明:il-4基因对带1～2 mm子叶柄的子叶的转化率显著高于下胚轴;带柄子叶在预培养1 d、侵染3 min、共培养1 d时,转化效果最好,转化率达23.3%;下胚轴在预培养1～3 d,侵染3～5 min,共培养1～2 d时,转化效果较好,转化率最高可达13.3%.经PCR检测及PCR-Southern杂交,初步证明目的基因il-4已经转入甘蓝的再生植株.转il-4基因甘蓝的PCR阳性再生植株已经开花并产生了后代.     

Screening and Identifying Two Specific Molecular Markers in Maintainer Line of Cytoplasmic Male Sterility Cauliflower （Brassica oleracea var. botrytis）

Cytoplasmic male sterility （CMS） is a maternally inherited trait that prevents the production of function pollen, but maintains female fertility. It has been widely used in breeding programs to product F1 hybrid seed in some crops （Hanson, 1991）. However, in some plants the CMS lines usually have some harmful characters such as plant dwarfness, etiolation in cauliflower. To obtain perfect CMS lines, a long time usually five to eight years is necessary. So how to fully utilize the obtained CMS lines for accelerating the development of new hybrid seed is very important. Here, in order to select high-quality paternal materials for cytoplasmic male sterility （CMS） cauliflower NKC-A in early stage, analyses of RAPD and ISSR were performed. Total 2 160 detectable bands were obtained by RAPD using 406 random primers. Among all these primers only the amplifications of primer S2121（5＇ GTGGCTCTCC 3＇） were polymorphic. A 934 bp band only existed in maintainer line. After cloning and sequencing, specific primers were designed to transform the RAPD marker into specific PCR marker, which was named S2121900. Furthermore, total of 30 ISSR primers were also analyzed which produced 306 detectable bands.     

Induction of an alloplasmic male-sterile Brassica oleracea by substituting cytoplasm from ‘Early Scarlet Globe’ radish (Raphanus sativus)

Summary Alloplasmic male-sterile Brassica oleracea L. was synthesized in a backcrossing program through amphidiploid Raphanobrassica by using Early Scarlet Globe radish (Raphanus sativus L.) as the donor of cytoplasm and B. oleracea broccoli and cabbage as recurrent pollen parents. Persistence of radish chromosomes and high female sterility were encountered in the first four backcrosses. Following use of colchiploid 4x broccoli as pollen parent, a BC₅ plant was obtained that had 2n=3x+1=28 chromosomes, improved seed set, and no radish traits. The BC₆ with recurrent 2x broccoli contained male-sterile plants with 2n=18 or 19 chromosomes, increased seed set, and broccoli morphology. Subsequent generations segregated for male-sterile and restored male-fertile plants, some with variable development of stamens and pollen. Leaf color of the alloplasmic plants, especially seedlings, was lighter green than normal.     

芸薹属物种(B.napus,B.rapa,B.oleracea)PHB基因的生物信息学分析

HD-ZIPⅢ基因家族参与植物胚胎发生、分生组织形成、叶极性建立和维管组织分化等过程。本研究利用生物信息学方法,对HD-ZIPⅢ成员之一PHABULOSA (PHB)进行鉴定,在甘蓝型油菜、白菜和甘蓝中共获得7个PHB成员,并对其核苷酸和氨基酸序列的基本特征、结构域、保守基序、基因结构和系统进化关系进行了分析。结果表明,这些PHB蛋白编码不稳定的亲水性蛋白,所有成员均含有Homeobox、bZIP、START和MEKHLA保守结构域,PHB位点在三个物种间的进化符合禹氏三角理论。序列分析及测序结果显示白菜的两个PHB基因在miR165、miR166结合位点处存在一个碱基差异。本研究为进一步揭示芸薹属物种PHB基因功能提供了依据。     

芸薹属物种(B.napus,B.rapa,B.oleracea)TCP基因家族全基因组鉴定和分析

TCP蛋白是植物特有的一类转录因子,广泛参与植物叶发育、花对称性、侧枝发育等过程,具有重要的调控作用。为了揭示芸薹属TCP基因家族的功能和进化关系,本研究利用生物信息学方法对白菜、甘蓝和甘蓝型油菜的TCP转录因子家族成员、基因分类、基因结构、系统进化关系和结构域序列保守性进行分析。研究表明,白菜和甘蓝的TCP基因成员各有37条、甘蓝型油菜TCP基因成员有78条,且全部为亲水蛋白,大部分的TCP基因结构简单,不含内含子;序列分析表明,这些TCP基因均具有高度保守的TCP结构域;根据结构域差异和系统发育分析的结果,将白菜、甘蓝、甘蓝型油菜TCP基因分为两类:ClassⅠ和ClassⅡ,ClassⅡ可继续分为CIN和CYC/TB1两个小组;聚类分析发现,芸薹属中白菜、甘蓝、甘蓝型油菜TCP基因与拟南芥垂直同源的At TCP11、At TCP16、At TCP23的TCP基因缺失,可能与芸薹属发生三倍化事件或驯化过程中的人工选择有关。以上研究将为揭示芸薹属TCP基因的功能提供重要的理论基础。     

芸薹属物种(B.napus,B.rapa,B.oleracea)CAMTA4基因家族的鉴定与进化分析

钙调蛋白在植物的生理代谢中起着重要作用,而钙调蛋白的转录翻译过程需要CAMTA的参与。为研究芸薹属植物中CAMTA4位点的分子进化,本研究利用生物信息学的方法,分析了甘蓝型油菜、白菜和甘蓝的CAMTA4家族核苷酸序列的基本特征和基因结构、蛋白序列的基本特征和功能域,以及它们的系统发生关系。结果显示,这3个物种的CAMTA4都具有CAMTA保守的结构域,且它们的进化符合禹氏三角理论和三倍化学说。本研究明确了甘蓝型油菜、白菜和甘蓝CAMTA4位点的进化历程,并且为研究芸薹属植物CAMTA的功能提供了生物信息学参考。     

芸薹属物种(B.napus,B.rapa,B.oleracea,B.juncea,B.nigra)MYB28家族的分子进化

芸薹属物种(甘蓝型油菜,白菜,甘蓝,芥菜型油菜和黑芥)是重要的经济作物,它们均含有硫代葡萄糖苷(简称硫苷),硫苷的合成受MYB转录因子的调控,其中MYB28能够调控甲硫氨酸合成脂肪族硫苷。目前,普遍认为芸薹属物种在进化过程中发生了全基因组三倍化,并且用禹氏三角表示来表示它们间的亲缘关系。本研究利用生物信息学方法,鉴定了5个芸薹属物种MYB28,分析了芸薹属MYB28的基本特征、同源性和系统发生关系,探讨了芸薹属物种MYB28家族的分子进化。结果显示,芸薹属MYB28在不同物种间的同源性存在高于种内同源性的现象,芸薹属MYB28位点的进化符合禹氏三角理论,但不完全符合三倍化学说。     

甘蓝型油菜(Brassica napus.L)种皮颜色与种子含油量的相关分析

通过光谱分析法定量测量了甘蓝型油菜(B.napus,L.)种子表面颜色的光度学和色度学指标,把甘蓝型油菜种皮颜色的变异分为8级,使相邻两级之间相差5个 NBS 色差单位。级别愈高,颜色愈深。通过相关和回归分析表明,在相同的遗传背景中,甘蓝型油菜种皮颜色与种籽含油量之间存在着显著负相关〔r=(-0.408)-(-0.662)〕。黄籽平     

甘蓝型油菜(Brassica napus.L)种皮颜色与种子含油量的相关分析

通过光谱分析法定量测量了甘蓝型油菜(B.napus,L.)种子表面颜色的光度学和色度学指标,把甘蓝型油菜种皮颜色的变异分为8级,使相邻两级之间相差5个 NBS 色差单位。级别愈高,颜色愈深。通过相关和回归分析表明,在相同的遗传背景中,甘蓝型油菜种皮颜色与种籽含油量之间存在着显著负相关〔r=(-0.408)-(-0.662)〕。黄籽平     

油菜(Brassica napus L.)矮化研究进展

油菜(Brassica napus L.)是具有重要影响的油料作物.通过杂交育种在油菜上的运用,获得了相较于旧品种在生活力、生长势、抗逆性、适应力、产量、品质等方面具有更大优势的新品种,但同时也带来了油菜倒伏与机械化生产率无法提高等问题.水稻和小麦相关的矮化研究与应用是绿色革命的重要组成部分,这为人们改良油菜品种提供了...     

Occurrence of metaxenia and false hybrids in Brassica juncea L. cv. Kikarashina × B. napus

Imported genetically modified (GM) canola (Brassica napus) is approved by Japanese law. Some GM canola varieties have been found around importation sites, and there is public concern that these may have any harmful effects on related species such as reduction of wild relatives. Because B. juncea is distributed throughout Japan and is known to be high crossability with B. napus, it is assumed to be a recipient of B. napus. However, there are few reports for introgression of cross-combination in B. juncea × B. napus. To assess crossability, we artificially pollinated B. juncea with B. napus. After harvesting a large number of progeny seeds, we observed false hybrids and metaxenia of seed coats. Seed coat color was classified into four categories and false hybrids were confirmed by morphological characteristics and random amplified polymorphic DNA (RAPD) markers. Furthermore, the occurrence of false hybrids was affected by varietal differences in B. napus, whereas that of metaxenia was related to hybridity. Therefore, we suggest that metaxenia can be used as a marker for hybrid identification in B. juncea L. cv. Kikarashina × B. napus. Our results suggest that hybrid productivity in B. juncea × B. napus should not be evaluated by only seed productivity, crossability ought to be assessed the detection of true hybrids.     

从油菜(Brassica napus L. cv. Topas)裂外壁小孢子培育出胚状体与小植株

油菜(Brassica napus L. Cv. Topas)小孢子经高温(32℃)预培养导致外壁开裂, 形成裂外壁小孢子. 微室饲养培养技术结合定位追踪观察证实裂外壁小孢子具有胚胎发生能力. 33.3%的裂外壁小孢子能启动细胞分裂, 其中13.3%的裂外壁小孢子能持续分裂, 并遵循胚胎发生途径. 与完整小孢子不同, 第一次分裂既有均等分裂, 亦有不均等分     

不结球白菜(Brassica campestris ssp. chinensis Makino)种质资源SRAP遗传分化分析

利用SRAP分子标记分析了国内外64份不结球白菜种质资源的DNA遗传多样性和遗传分化。21对引物组合共检测出215个位点,其中112个为多态性位点,多态性比率达52.09%,平均每对引物组合产生10.24个位点和5.33个多态性位点。不结球白菜各类型中普通白菜的Nei’s基因多样性指数（0.1410）和遗传丰富度[(190) 88.37%]最高;各生态区域中江淮流域不结球白菜的Nei’s基因多样性指数（0.1451）和遗传丰富度[(185) 86.05%]最高;国内的Nei’s基因多样性指数(0.1293)和遗传丰富度[(188) 87.44%]分别高于国外。遗传变异估算表明,不结球白菜遗传分化系数58.22%,大部分变异存在于种群间;基因流为0.4031,说明群体间基因流动较少,遗传分化程度较高。以遗传相似系数0.872为截值,可把不结球球白菜分为Ⅵ个类群。