细胞分裂素对甘蓝型油菜种皮色泽形成的影响

以3对遗传背景相同的甘蓝型黄籽和黑籽油菜为材料,研究甘蓝型油菜种子发育过程中內源细胞分裂素(ZR)、各种色素、色素合成相关酶活性的动态变化及其相互关系,并以外源细胞分裂素类物质(6-BA)加以验证,结果表明,相同遗传背景下的黄籽油菜种子的ZR含量较黑籽油菜高,花后27 d比黑籽高4~5倍; 在甘蓝型黄籽油菜种子发育前期(27 d阶段)种子中细胞分裂素含量越高其成熟种子色泽就越浅; 种子的ZR含量与种皮中类黄酮、花色素、黑色素含量显著负相关,与多酚含量显著正相关,与酪氨酸酶显著负相关,与苯丙氨酸解氨酶、多酚氧化酶无显著相关性; 施用外源细胞分裂素6-BA (50 mg L^–1)可显著提高黄籽油菜黄籽度,明显降低甘蓝型油菜种皮中黑色素、花色素、类黄酮含量,对黑籽种皮的多酚含量无显著影响,但可增加黄籽种皮多酚含量; 6-BA处理可降低油菜种皮中酪氨酸酶、苯丙氨酸解氨酶活性,对多酚氧化酶活性无显著影响。表明细胞分裂素可减缓甘蓝型油菜种皮各色素合成,从而影响黄籽油菜色泽;该过程可能是通过调控色素合成的相关酶活性来实现的。     

芸苔属主要油料作物黄籽性状分子遗传研究进展

油菜黄籽性状由于较好的营养和加工品质,得到现代油菜育种越来越多的重视,正逐渐成为油菜育种的重要目标.本文主要从芸苔属3个主要油料作物黄籽性状的遗传规律、基因定位和比较基因组学研究进展进行综述.在黄籽性状遗传方面,白菜型油菜、芥菜型油菜的遗传模式己基本清楚,由两对隐性基因控制,而甘蓝型油菜由于黄籽基因来源不同,存在多种遗传模式.在黄籽基因的定位方面,白菜型油菜、芥菜型油菜均已筛选到多个与黄籽性状紧密连锁的分子标记,并成功转成适合大规模筛选的SCAR标记,为分子标记辅助育种奠定了基础.甘蓝型油菜中也获得一些与黄籽性状紧密连锁的分子标记,并应用于分子标记辅助育种,但由于黄籽来源的不同,这些标记缺少通用性.在比较基因组学方面,根据拟南芥的相关种皮颜色基因,在白菜型油菜、芥菜型油菜、甘蓝型油菜中已克隆出一些与种皮颜色合成有关的基因,这些基因的克隆为油菜色素合成途径和通过转基因创造新型黄籽油菜研究奠定了基础.     

甘蓝型黄籽油菜的遗传研究

从1975—1990年作者及其研究集体系统坚持了甘蓝型黄籽油菜的遗传育种研究。15年来取得了以下主要研究结果: 1.甘蓝型黄籽油菜的种皮色泽不同于白菜型、芥菜型和埃塞俄比亚油菜,为土黄或姜黄,而没有纯黄,只有杂黄,即在黄色种皮上有黑色斑点、斑块或褐色环带。 2.长期自交后仍得不到遗传上稳定的纯黄后代;在大群体中,不论是自     

白菜型油菜黄籽遗传研究

利用4个白菜型油菜品种进行黄籽与非黄籽之间的正反交和相应的回交测交,以探讨其种皮颜色的遗传及其与其它性状的关系。结果表明:白菜型油菜种皮颜色具有母体效应。黄籽受两对重叠隐性基因控制。在黄籽与黑籽之间存在两对显隐性关系,而在黄籽与褐籽之间只存在一对显隐性关系。在相同遗传背景下,黄籽含油量比非黄籽要高,皮壳率要低,而千粒重差异不显著。     

不同环境中甘蓝型油菜种皮木质素含量的QTL定位

以甘蓝型黄籽油菜GH06和甘蓝型黑籽油菜中油821为亲本杂交,后代通过“一粒传法”连续自交7代构建重组自交系,2007年分别在重庆市北碚区和万州区2个试验基地种植重组自交系群体,利用本实验室已构建的遗传连锁图谱和复合区间作图法(CIM),对种皮木质素含量进行测定及QTL定位分析。结果在2个环境中共检测到12个种皮木质素含量相关的QTL,分别位于4个不同的连锁群,单个QTL可解释性状表型变异的4.50%~8.79%;在第3连锁群上检测到1个QTL与同一标记EM19ME23/130连锁,其余10个QTL位置不同;在北碚检测到的QTL主要分布于第20连锁群,在万州检测到的QTL主要位于第3连锁群;部分种皮木质素的QTL与种胚类黄酮和种皮色泽的QTL位于相近区间,在北碚和万州种皮木质素含量与种胚类黄酮存在极显著和显著正相关关系。结果表明甘蓝型油菜种皮木质素含量表现为多基因控制的数量性状,基因表达受环境影响较大;油菜种皮木质素合成和类黄酮的积累可能受相同关键基因调控或者具有部分相同的合成代谢途径。     

甘蓝型纯黄籽油菜新品系YR 5602的选育

通过甘白远缘杂交和复合杂交相结合的方法，经7年8代选育，于2004年育成了甘蓝型纯黄籽双低油菜新品系YR5602，其芥酸含量为1．18％～1．66％，硫甙28．8～38．3／μmol／g，含油量43．8％～45，2％，种子蛋白质23．3％～24．6％；群体黄籽株率达100％，黄籽频率达100％，黄籽度高达1．94～2．0，除种脐颜色稍深外，种皮全部为淡黄色且无斑点，不论是自交种或是天然种，其黄籽性状均表现极为稳定。已有两个甘蓝型油菜纯黄籽双低组合进入贵州省预备试验。至目前为止，象这样稳定的纯黄籽甘蓝型油菜双低高油分材料极为少见，是1份极为珍贵的重要种质资源，对甘蓝型油菜黄籽优质育种与杂种优势利用具有很高的利用价值．     

甘蓝型黄籽油菜主要品质性状间的相关性分析

【研究目的】为探讨不同田间环境条件下,同一甘蓝型黄籽油菜基因型各品质性状之间的相关关系,【方法】以甘蓝型黄籽油菜基因型GH01为材料,研究了由播期与密度形成的9种田间环境条件下粒色及其它主要品质性状间的相关性。【结果】结果表明：黄籽度与胚含油量、种子含油量间呈明显的负相关关系（相关系数分别为r=-0.6617和r=-0.7458*）,胚含油量与胚蛋白质含量、种子含油量之间仍保持着极其显著的负相关和正相关关系（相关系数分别为r=-0.9071**和r=0.8973**）。【结论】甘蓝型黄籽油菜黄籽度、种子含油量之间的相关关系随供试基因型的不同而发生变化,但另一些品质性状（含油量、蛋白质含量）间的相关关系并不因供试基因型间的差异而改变。     

甘蓝型黄籽油菜主要品质性状与环境效应 基因型差异研究

以8个不同来源的甘蓝型黄籽油菜基因型为材料,研究了4种不同密、肥(氮素)处理条件下粒色及其它主要品质性状的变化。结果表明:不同来源的甘蓝型黄籽油菜基因型,其粒色等品质性状存在极显著差异;甘蓝型黄籽油菜的品质性状主要决定于基因遗传效应,同时密肥效应和基因型与密肥的互作效应对甘蓝型黄籽油菜的品质也有极显著影响。不同基因型甘蓝型黄籽油菜的品质性状在密肥处理间变异程度不同。     

甘蓝型黄籽油菜性状组间的典型相关分析

以7个不同遗传来源的甘蓝型黄籽品系所配制的完全双列杂交作样本,对甘蓝型黄籽油菜进行典型相关分析。结果表明,在性状组间关联上起主要作用的是株型性状中的株高、产量性状中的单株粒重、品质性状中的皮壳率和黄籽度。用多目标综合选择指数,讨论了甘蓝型黄籽油菜的选择问题。     

甘蓝型纯黄籽油菜YR 5602种子粒色遗传研究

经遗传研究证明,甘蓝型纯黄籽油菜YR 5602种子粒色为1对隐性基因控制,种皮颜色受母本基因型控制,不受花粉和胚基因的影响,种子粒色能稳定遗传。     

Phenotypic variation in progenies from somatic hybrids between Brassica napus and Sinapis alba

A number of variant phenotypes, such as long siliques, high density of pods, increased seed number per pod and yellow seed color resembling Sinapis alba, were selected from intergeneric somatic hybrids between Brassica napus and Sinapis alba through successive backcrosses. Resistance to Sclerotinia sclerotiorum among 24 BCF₄ lines was also tested by inoculation with mycelia, and four lines showed stronger resistance than the resistant rapeseed variety ‘Zhongshuang 9’. Comparative anatomy studies on mature seed revealed that the seed coat pigments were mainly distributed in the palisade layer, which is considerably thinner in S. alba compared to B. napus. The area index of protein bodies in cotyledon cells was highest in S. alba, lowest in B. napus and intermediate in the intergeneric progenies. This study demonstrates that wide hybridization can enable exploitation of valuable trait diversity from S. alba for broadening the genetic basis for rapeseed breeding.     

Inheritance of resistance to the papaya ringspot virus-watermelon strain from two different accessions of winter squash Cucurbita maxima Duch.

Researchers have conclusively shown that Sinapis alba (commonly known as yellow mustard) has many agronomic traits which would be beneficial if transferred to rapeseed ( Brassica napus L.). S. alba is resistant or tolerant to all major insect pests of Brassica crops in the Pacific Northwest region of the United States of America. It is also tolerant of high temperatures and drought stress, is shatter resistant and capable of high seed yield without the need for insecticides and herbicides. However, S. alba is considerably lower in oil content and lacks the high oil quality and seed meal quality of rapeseed (i.e. canola). This paper describes a combination of ovary culture and embryo rescue techniques used to develop fertile hybrid plants from the intergeneric cross between S. alba and B. napus . The hybrids were intermediate between both parents for presence of trichomes, leaf shape and color, seed size, pod shape, and seed oil content; showing expression of traits from both parental species. Hybrid plant tissue and seed contained all types of glucosinolate that exists in either B. napus or S. alba, at the same or higher level to the parental species. These hybrid crosses offer the potential for combining the desirable oil and glucosinolate qualities of B. napus with insect and disease resistance characters of S. alba.     

黄黑籽甘蓝型油菜类黄酮途径基因SNP位点检测分析

类黄酮物质在植物花、叶、果实和种子颜色变化的过程中起着至关重要的作用,本研究以不同黄黑籽种皮材料为研究对象,采用基因同源克隆方法,获得17个类黄酮基因全长ORF序列,在核酸和蛋白水平上分别序列差异比较表明,这些基因在不同黄黑籽材料中共存在41个不同拷贝成员。在核苷酸水平上,检测到BnTT3、BnTT18、BnTTG1和BnTTG2的单核苷酸位点数目介于16~52之间,且BnTTG2在3个不同的位置上还存在多个碱基的连续性缺失现象(119~121 bp,183~189 bp和325~330 bp),但在蛋白水平上仅存在2~16个氨基酸位点差异,说明BnTT3、BnTT18、BnTTG1和BnTTG2在不同甘蓝型黄黑籽材料中存在单核苷酸位点差异,而单核苷酸位点突变不一定导致氨基酸位点的变异。在不同黄黑籽材料中仅BnTT3和BnTT18存在一致性的氨基酸突变位点(252和87),推测BnTT3和BnTT18可能在黄黑籽甘蓝型油菜种皮颜色差异形成过程中发挥至关重要的作用。通过这些位点的等位特异PCR可以区分材料间透明种皮基因,为特异基因芯片的开发及阐明甘蓝型油菜种皮色泽性状的基因及其作用位点奠定基础。     

甘蓝型油菜BnTT3基因的表达与eQTL定位分析

类黄酮途径中,TT3编码的4-二氢黄铜醇还原酶是参与原花色素和花青素合成的关键酶。为了明确该基因可能的上游调控网络,利用黄籽母本GH06和黑籽父本ZY821构建的遗传图谱,以Bn TT3基因在高世代重组自交系群体中随机选取的94个株系花后40 d种子的表达量作为性状,采用复合区间作图法进行e QTL分析。结果共检测到5个表达量相关的e QTL,分别位于A03、A08、A09和C01染色体,单个e QTL解释表型变异的5.22%~24.05%。A09染色体上存在2个主效e QTL,单个e QTL分别解释24.05%和16.55%的表型变异,分别位于标记KS10260~KBr B019I24.15和B055B21-5~KS30880之间,微效e QTL分布于A03、A08和C01染色体上。A09染色体上的2个主效e QTL区间(包含200 kb侧翼序列)与拟南芥、白菜、甘蓝和芸薹族近缘物种基因组同源区段具有很好的共线性关系。基因注释结果表明检测到的e QTL均为trans-QTL,2个主效e QTL区段共包含78个基因,包括MYB51、MYB52和b ZIP5转录因子,可能为Bn TT3基因的上游直接调控因子,对这些基因功能的深入分析将有助于阐明甘蓝型油菜黄籽性状形成的分子调控机制,为黄籽候选基因的克隆筛选奠定基础。     

Persistent C genome chromosome regions identified by SSR analysis in backcross progenies between Brassica juncea and B. napus

Given that feral transgenic canola (Brassica napus) from spilled seeds has been found outside of farmer’s fields and that B. juncea is distributed worldwide, it is possible that introgression to B. juncea from B. napus has occurred. To investigate such introgression, we characterized the persistence of B. napus C genome chromosome (C-chromosome) regions in backcross progenies by B. napus C-chromosome specific simple sequence repeat (SSR) markers. We produced backcross progenies from B. juncea and F₁ hybrid of B. juncea × B. napus to evaluate persistence of C-chromosome region, and screened 83 markers from a set of reported C-chromosome specific SSR markers. Eighty-five percent of the SSR markers were deleted in the BC₁ obtained from B. juncea × F₁ hybrid, and this BC₁ exhibited a plant type like that of B. juncea. Most markers were deleted in BC₂ and BC₃ plants, with only two markers persisting in the BC₃. These results indicate a small possibility of persistence of C-chromosome regions in our backcross progenies. Knowledge about the persistence of B. napus C-chromosome regions in backcross progenies may contribute to shed light on gene introgression.     

甘蓝型油菜与播娘蒿原生质体融合杂种后代的遗传研究

甘蓝型油菜(2n=38)与播娘蒿(2n=28)原生质体融合杂种F₁连续自交3代,获得F₂、F₃和F₄后代。用细胞学和SSR分子标记方法,分析杂种后代的染色体数目变异、减数分裂行为以及播娘蒿遗传成分的保留情况。结果表明在F₂、F₃和F₄代中,根尖细胞染色体平均数分别为38.47±3.17、37.65±3.23和36.66±2.95,随着自交世代增加呈减少趋势;在杂种后代减数分裂中,观察到染色体桥、染色体落后、染色体周期不同步、不均等分离等现象;杂种后代F₂、F₃和F₄代中检测到播娘蒿特征条带的平均频率分别为9.62%、2.99%和0.31%,呈减少趋势。因此要实现播娘蒿种质向油菜渗入应该重视F₂世代的选择。     

Phenotypic and seed structural comparison in hybrids of different Brassica rapa and B. oleracea

Brassica napus is an important oil species with short history and narrow genetic background. Interspecific hybrids from crosses between B. oleracea and different B. rapa were obtained. We found the hybrids with white petal resembling B. oleracea, the flavonoid and phenolic content decreased in hybrids, agreeing with the expressional changes of flavonoid biosynthesis genes. Seed coat of hybrids resembled diploid parents, or partly resembled to each parent with a clear outline. The palisade layer in hybrids was thicker than parents, with similar pigment accumulation as B. oleracea but more than B. rapa. Differentially sized protein bodies (PBs) were found in hybrids. The radical and inner cotyledon of all hybrids were identified with larger but less PBs than parents. The average size of PBs in outer cotyledon of resynthesized B. napus was also larger than parents, but the number of PBs was not significantly reduced. The phenotypic and seed structural variations after polyploidization of B. napus would be interesting for genetic broadening and breeding of rapeseed.     

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.     

芥菜型油菜黄籽性状的遗传、基因定位和起源探讨

油菜种皮颜色既是一个形态指示性状, 又与种子休眠和品质有关。以芥菜型油菜种皮颜色分离的2个BC₆F₂群体为作图群体,用微卫星(SSR)等标记进行连锁定位, 并用定位标记对22份材料进行关联分析, 通过反转录-聚合酶链反应(RT-PCR)分析12份材料种皮中4-二氢黄酮醇还原酶(DFR)、花色素合酶(ANS)和花色素还原酶(ANR)基因的表达, 对6份黄籽材料的种皮颜色基因等位性进行测定, 结果将芥菜型油菜控制种皮颜色的2个基因位点分别定位到A9和B3连锁群, 并找到其两侧紧密连锁标记, 发现黄籽材料种皮颜色基因位点附近0.9 cM和1.5 cM区域高度保守, 所有黑色种皮中DFR、ANS和ANR基因均表达, 所有黄色种皮中DFR和ANS均不表达,但ANR基因表达或不表达,黄籽材料的种皮颜色基因等位。根据这些结果结合前人研究, 认为芥菜型油菜种皮颜色基因是调控基因,黄籽为单一起源。     

甘蓝型油菜种皮花青素相对含量测定方法研究

在配合甘蓝型油菜育种工作中,花青素含量的变化是重要的一个测定指标,笔者介绍了一种改进的测定甘蓝型油菜种皮中花青素相对含量的方法pH差示法,同常用的盐酸甲醇法相比较,同样不需要标准样品,成本低廉,测定简便快捷,同时能有效的去除甘蓝型油菜种皮中褐色物质对测定结果的干扰,所得结果能更准确地表现出不同色泽的种皮中花青素含量的变化。pH差示法在简便,快捷的基础上,所测定甘蓝型油菜种皮花青素相对含量更为准确、有效。