Sexual hybridisation in crosses of cultivated Brassica species with the crucifers Erucastrum gallicum and Raphanus raphanistrum: Potential for gene introgression

Studies were conducted to investigate the crossability of the cultivated Brassica species, Brassica napus (oilseed rape), B. rapa (turnip rape), and B. juncea (brown and oriental mustard), with two related cruciferous weeds that are abundant in certain regions of Canada, Erucastrum gallicum (dog mustard) and Raphanus raphanistrum ssp. raphanistrum (wild radish). Seed was produced without recourse to embryo rescue from all reciprocal crosses except R. raphanistrum × B. juncea. Four hybrid plants were recovered, namely B. napus × E. gallicum, B. napus × R. raphanistrum (two plants), and B. rapa × E. gallicum. The hybrids were characterized by their morphology, RAPD analysis, and cytological examination. The B. rapa × E. gallicum hybrid was extremely vigourous and fertile, and would likely grow in natural habitats. This hybrid produced self-seed and backcrossed readily with the B. rapa parent and, to a lesser extent, with the E. gallicum parent. The B. napus × E. gallicum hybrid was a weak plant, but produced fertile backcross progeny with the E. gallicum parent. The B. napus × R. raphanistrum hybrids were vigourous but mostly sterile. Because of their low vigour and/or sterility, hybrids produced from crosses of B. napus with the cruciferous weeds would not likely be an environmental concern. However, the potential of the B. napus × E. gallicum and B. rapa × E. gallicum hybrids to backcross with E. gallicum may be of concern. This revised version was published online in July 2006 with corrections to the Cover Date.     

甘蓝型油菜与近缘种、属杂交时花粉-雌蕊相互作用的研究

甘蓝型油菜作母本与白菜、甘蓝、黑芥、芥菜型油菜和埃塞俄比亚芥杂交时,花粉粘合程度减弱。花粉管常常不能穿入柱头,与花粉管接触的柱头乳突细胞内普遍产生胼胝质。少数异源花粉管能穿过柱头进入花柱,但时有异常胼胝质沉积在花粉管中。海甘蓝的花粉很难粘合在甘蓝型油菜柱头上。异种花粉与甘蓝型油菜雌蕊的亲和性按大小依次     

油菜及其近缘种种子萌发期耐盐碱性差异

为了明确甘蓝型油菜、白菜型油菜、芥菜型油菜、白芥和芸芥种子萌发期的耐盐碱性差异,分别采用0（CK）、0.4%、0.6%、0.8%、1.0%、1.2%和1.5%的NaCl溶液和0（CK）、0.1%、0.2%、0.3%、0.4%、0.5%和0.6%的NaHCO₃溶液进行萌发期胁迫处理,测定了发芽势、发芽率和发芽指数的变化规律。结果表明：不同浓度NaCl溶液处理,油菜及其近缘植物的平均发芽率依次为芥菜型油菜（71.7%）>甘蓝型油菜（64.0%）>白芥（62.4%）>白菜型油菜（48.4%）>芸芥（30.6%）。当NaCl的浓度为1.0%时,芥菜型油菜的发芽率最高,为91%,芸芥最低,仅有2%。不同浓度NaHCO₃溶液胁迫下,5种植物的平均发芽率依次为白芥（85.3%）>甘蓝型油菜（84.3%）>白菜型油菜（59.1%）>芸芥（42.9%）>芥菜型油菜（34.3%）。当NaHCO₃溶液浓度为0.4%时,白芥的发芽率最高,为95%,而芥菜型油菜最低,为0。综上,芥菜型油菜对NaCl胁迫的耐性最强,白菜型油菜和芸芥对NaCl胁迫的耐性差;白芥和甘蓝型油菜对NaHCO₃胁迫的耐性强,而芥菜型油菜对NaHCO₃胁迫的耐性较差。可见,在油菜种植区改良盐渍化土壤,中性盐渍化土壤最好选用芥菜型油菜,偏碱性盐渍化土壤可以选用白芥或甘蓝型油菜。     

Cytoplasm Influences the Photosynthetic Efficiency in Brassica carinata

Cytoplasmic effects on the photosynthesis was investigated in experimentally produced Brassica carinatas of reciprocal cytoplasmic origin. These are obtained by hybridizing B. nigra x B. oleracea var. italica ( B. carinata 226) and B. oleracea x B. nigra ( B. carinata 241). Natural B. carinata and the two parents viz. B. nigra and B. oleracea var. italica were also included in this study. It was observed that the rate of photosynthesis in B. carinata 226 with nigra cytoplasm, and natural B. carinata was higher than that of B. carinata 241 with oleracea cytoplasm. This was substantiated by the diurnal values of photosynthesis among these cultivars. The result indicated that the cytoplasm does affect the rate of photosynthesis in B. carinata and high photosynthetic efficiency in natural B. carinata was due to B. nigra maternal parent.     

Macro-nutrient Status of Brassica napus and Brassica juncea Grown Under Swedish Conditions

Concentrations and amounts of macro-nutrients (N, P, K, Ca, Mg, S) in shoots, vegetative and generative plant parts in B. napus and B. juncea and nutrient inflow per unit root length in B. napus were studied in a field experiment at Uppsala, Sweden. Concentrations in vegetative plant parts were, except for Ca in B. napus, highest at the beginning of the season, and N and K were higher in B. napus than in B. juncea. The higher N concentration in vegetative parts (leaves, stems and pods) in B. napus was probably due to differences in growth pattern between the species. Seed concentrations of N, P, Ca, Mg and, most pronounced of all, S were higher in B. juncea. Nutrient amounts were, with the exception of K, throughout the season higher in B. juncea than in B. napus due to a higher dry matter production. Nutrient inflow per unit root length in B. napus was highest during the rosette stage for all nutrients. During flowering, the inflow decreased for N and K and increased for P and Ca. After root growth had stopped, no net inflow of K and S was found and inflow rates of other nutrients were at very low levels.     

Contribution of Leaf Photosynthesis towards Seed Yield in Brassica species

The effect of defoliation after flowering, when a considerable number of pods were already formed on yield components, was examined in four Brassica species. In B. campestris and B. juncea , a decrease in seed yield by defoliation was associated with a decrease in the number of pods per plant as seed number per pod and thousand-seed weight were not significantly affected by this treatment. In B. nigra , apart from decreasing the pod number per plant, defoliation also significantly decreased the thousand-seed weight, Defoliation significantly decreased both seed number per pod and thousand-seed weight apart from decreasing the number of pods per plant in B. carinata. The study indicated that in B. campestris and B. juncea , the role of leaf photosynthesis seemed to be restricted mainly to the formation of pods. On the other hand, in B. nigra and B. carinata it appeared that leaf photosynthesis also contributed towards seed development.     

芸芥(Eruca sativa Mill.)与芸薹属(Brassica L.) 3个油用种的远缘杂交

采用芸芥(Eruca sativa Mill.)与芸薹属3个油用种（Brassica napus, Brassica juncea, Brassica rapa）进行杂交，共授粉15 990朵花，获得1 257个角果，711粒杂交种子，结角率为7.86%，亲和指数0.045。经形态学鉴定，无论芸芥作母本还是芸薹属的三个油用种作母本，F1植株均为偏母植株。杂交所获得角果的角粒数很低，许多角果     

Brassilexin accumulation and resistance to Leptosphaeria maculans in Brassica spp. and progeny of an interspecific cross B. juncea × B. napus

Summary Resistance to Leptosphaeria maculans was assessed in Brassica napus, B. juncea, B. carinata, B. nigra and progeny issuing from an interspecific cross B. napus × B. juncea, using a cotyledon-inoculation test. In these individual plants, brassilexin accumulation was determined following an abiotic, non-specific, elicitation. All the tested B. napus cultivars were highly susceptible to the parasite and weakly accumulated brassilexin. In contrast, B. juncea, B. carinata, and B. nigra usually displayed a hypersensitive response to the inoculation and accumulated more brassilexin than B. napus. The same correlation between resistance to L. maculans and phytoalexin accumulation was observed in the interspecific hybrid progeny. The cotyledon-inoculation test allowed the discrimination of plants displaying a hypersensitive response to the inoculation from those highly sensitive to the parasite, but intermediate disease severity classes were not usually representative of resistance or susceptibility. In this respect, brassilexin determination allowed differentiation, within a set of plants presenting an intermediate response to the pathogen, of plants with a high (B. juncea-like), and with a weak (B. napus-like) ability to accumulate brassilexin.Abbreviations IHP interspecific hybrid progeny - JR B. juncea-type complete resistance to blackleg (Roy, 1984) - W&D test cotyledon-inoculation test as described by Williams & Delwiche (1979)     

Genetic diversity of Brassica carinata with emphasis on the interspecific crossability with B. rapa

Brassica carinata (BBCC, 2n = 34) possesses many good agronomic characters and, to provide more genetic information about this species and utilize it further, 110 accessions of B. carinata were tested for genetic variation by using 233 amplified fragment length polymorphism markers, so that a dendrogram could be constructed. To combine the good traits of B. carinata and B. rapa (AA, 2n = 20) with those of B. napus (AACC, 2n = 38) which is the major rapeseed variety in China, interspecific crosses between these two species have been made which have resulted in 276 doubled hybrids (AABBCC). These hexaploids combine positive characters and can be used for crossing with B. napus . Brassica carinata as the female parents exhibited extensive differences in interspecific crossability and 20 high crossability accessions were identified. The Poisson Regression Model analysis result (P < 0.0255) indicated that such differences were due to the genotype of the accessions. Accessions with high crossability could promote gene flow within the genus Brassica .     

The production of yellow-seeded Brassica napus (AACC) through crossing interspecific hybrids of B. campestris (AA) and B. carinata (BBCC) with B. napus

To transfer the genes for yellow seed coat from both genomes A and C to B. napus (AACC), the hexaploid of Brassica (AABBCC) was synthesised from reciprocal interspecific crosses between yellow-seeded B.campestris (AA) and B.carinata (BBCC). The hexaploid with 27 pairs of chromosomes was red-seeded which showed that genic interaction existed in the trigenomic plants for the colour of the seed coat. Hundreds of hybrid seeds were obtained from crosses between the red-seeded hexaploid and partial yellow or brown-seeded varieties of B. napus as pollen donor. The majority of the hybrid plants (AABCC) were self fertile with brown seeds. It appeared that the chromosomes of the B genome were excluded during the meiosis of the pentaploid and a high proportion of the genetically balanced AC gametes could be produced. The fertility of the F2 population was increased and even reached normal levels for some plants. Seventy-three plants with the yellow-seeded character were isolated from 2590 open-pollinated F2 plants, most with increased fertility. After two successive self-pollinations, 18 lines produced yellow seeds and no brown seeds segregated from these populations. The morphology of the novel yellow-seeded plants was basically towards B. napus. Esterase isoenzyme electrophoresis showed that the plants contained some of the genetic background of B. campestris, B. carinata and B. napus. Cytological analysis has shown that at least some yellow-seeded lines have the B.napus AACC genome composition with 38 chromosomes and normal meiotic pairing. This revised version was published online in July 2006 with corrections to the Cover Date.     

芥菜型油菜黄黑种皮多酚差异的紫外-可见光谱研究

以芥菜型自交系黄黑油菜种皮为材料,研究了其多酚提取液的紫外-可见吸收光谱, 测定了芥菜型油菜黄黑种皮样以没食子酸和茶儿茶素标准计的多酚含量,发现黄种皮的多酚含量约为黑种皮的90%,显著区别于甘蓝型油菜的10%,因此多酚含量高可能是芥菜型黄籽油菜性状稳定的根本内因。基于香草醛试验,推测黑种皮中含有3-OH的黄烷醇类多酚,而黄种皮中没有。基于加入位移试剂后特征吸收谱带Ⅰ与Ⅱ的位移方向与大小,推测黄酮醇类多酚A环在黄种皮中可能主要为单酚羟基,而在黑种皮中可能主要为邻二酚羟基结构。这种多酚分子的结构差异可能是形成芥菜型油菜黄黑种皮色泽差异的主要原因。     

芥菜型油菜与甘蓝型油菜嫁接嵌合体的性状表现

以芥菜型油菜(B. juncea)作砧木,以甘蓝型油菜(B. napus)作接穗,形成嫁接嵌合体（以下简称“甘+芥”嫁接嵌合体）,对其植物学性状、农艺性状和32P标记的难溶性磷矿粉的吸收能力进行了研究。结果表明,“甘+芥”嫁接嵌合体除叶片出现紫色性状外,其他性状与甘蓝型油菜没有差异,但其农艺性状明显优于甘蓝型油菜和芥菜型油菜。其根系对肥料中难溶性磷吸收能力强,而且多数转移到地上部分,在体内分布较合理。     

几种芸薹属油菜的起源和进化

本文主要介绍油菜的起源和进化情况,特别是有关中国芸薹和芥菜型油菜,以及从国外引进的甘兰型油菜的历史考证。近年来,由于在华中和西北发现山芥菜和野生型黑芥,可以得出结论说,油菜是多源发生的,中国是白菜型油菜和芥菜型油菜的发源中心之一。白菜型原始祖先 B.compostris 起源于华中和西北,B.chinensis 型油菜起源于中国中     

Interactive Effects of Elevated CO2 and Moisture Stress on the Photosynthesis, Water Relation and Growth of Brassica Species

Interactive effects of elevated CO2 and moisture stress on photosynthesis, growth and water relation of Brassica species were studied using open top chamber technology. Brassica species responded to the elevated CO2 significantly under moisture stress condition. The adverse effect of moisture stress on the photosynthesis and plant water components were minimized by elevated levels of CO2. Drought susceptible species of B. campestris and B. nigra responded better to elevated CO2 compared to drought tolerant Brassica species such as B. carinata and B. juncea. The plant water potential significantly improved by elevated CO2 coupled with higher stomatal resistance and root growth.     

转基因甘蓝型油菜向其近缘种的基因漂移之研究进展

从甘蓝型油菜的生活周期出发,概述了甘蓝型油菜及其近缘种的开花习性、甘蓝型油菜与其近缘种的杂交亲和性、杂种及其回交后代的育性和适应性,评价了在中国环境条件下甘蓝型油菜中的外源基因向其近缘种漂移而导致生态风险的可能性。认为在中国环境条件下,转基因甘蓝型油菜向其野生杂草近缘种的基因漂移风险较小,而如何防止转基因向白菜型油菜（Brassica rapa）和芥菜型油菜（B.juncea）转移应作为中国转基因甘蓝型油菜基因基因漂移风险控制的关键。     

Effect of Elevated CO2 on the Photosynthesis, Growth and Water Relation of Brassica Species Under Moisture Stress

An attempt has been made to study the interactive effect of elevated CO₂ and moisture stress on photosynthesis, growth and water relations of Brassica species using open top chambers. It was observed that plants responded to elevated CO₂ significantly under moisture stress condition mitigating the adverse effects on photosynthesis and growth of Brassica species. Relatively drought susceptible species, viz. B. campestris and B. nigra , responded to elevated CC₂ markedly as compared to less sensitive B. carinata and B. juncea plants. The water status of plants significantly improved under elevated CO₂ concentration possibly by increasing stomatal resistance and/or by increased root growth.     

甘蓝型黄籽油菜遗传分类及遗传多样性研究

黄籽油菜的遗传研究是实现油菜籽粒高含油量且饼粕低纤维素、低单宁、低色素等育种目标的重要途径。为了对不同来源的黄籽油菜进行遗传分类和多样性研究,对黄籽材料进行遗传等位性测验及聚类分析。结果表明,经过遗传测验,按照相互杂交F1、F2粒色是否分离将11份黄籽材料分为5组,‘油研10’、CZV55、E718、Arm为一组,为一般显性遗传;Q33、D615为一组,为不完全显性遗传;2006C、X2006、740C为一组;Polo为一组;HY15为一组,后3组为一般隐性遗传。但是,2006C、‘油研10’等的黄籽性状的遗传根据杂交测验亲本的不同而呈现显性或隐性的变化。依据SSR标记检测结果进行聚类分析,将35份黄籽材料分为甘蓝型油菜、白菜型、芥菜型三大组。甘蓝型油菜又可再分为8个亚组,各自代表材料有2006C和740C、油研系统、Q33;法国Ramiro、陕西的GQ4、源自加拿大的Arm、prof;源自波兰的Polo等。不同材料按照系谱关系、育种单位、地理来源聚集在一起。研究结果为黄籽油菜杂种优势利用奠定理论基础。     

甘蓝型油菜对华北坝上冷凉环境的适应性

针对华北坝上冷凉区短季类经济作物缺乏,芥菜型油菜品质差、产量低等问题,于2017-2018年在农业农村部张北农业资源与生态环境重点野外观测试验站,引种了甘蓝型双低油菜品种华油杂62,以当地传统芥菜型油菜品种大黄为对照,监测了两品种的叶面积指数、角果净光合速率日变化、地上部干物质积累量、养分积累量和油菜农艺性状、产量等。结果表明,华油杂62较大黄生育期长约10d,平均叶面积指数高23.36%~36.73%;角果净光合速率不及大黄,N、P、K积累量高于大黄;干物质快速增长期延长21d,干物质积累潜值高46.86%;菜子产量较大黄高22.8%~54.0%。甘蓝型油菜华油杂62对华北坝上冷凉区的地理气候环境有较好的生态适应性,具有替代区域传统芥菜型油菜的优势。     

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MYB4基因RNA干扰载体构建

拟南芥(Arabidopsis thaliana)MYB4(AtMYB4)编码负调控转录因子,通过对苯丙烷途径中关键酶C4H(肉桂酸4-羟化酶)基因的抑制来调控重要次生代谢物质芥子酸酯的生物合成,在防御紫外线中起重要作用。同为十字花科的芸薹属(Brassica)有多种重要的油料、蔬菜和园林作物,如甘蓝型油菜(B.napus)、白菜(B.rapa)、甘蓝(B.oleracea)和芥菜(B.juncea)等。本研究克隆了甘蓝型油菜MYB4基因家族的RNA干扰片段BMYB4I,将其反义片段、正义片段采用NcoI+AatII和BamHI+XbaI分别插入到改进型植物RNA干扰基础载体pFGC5941M的启动子与间隔区、间隔区与终止子之间,形成重组载体pF-GC5941M-BMYB4I(简称为pBMYB4I)。经复合PCR鉴定表明,载体构建成功,并转化根癌农杆菌获得工程菌株。为揭示芸薹属MYB4基因调控芥子酸酯等苯丙烷物质合成的分子机理以及探索抗紫外线分子育种奠定了基础。