芥菜型油菜与埃塞俄比亚芥杂种基因组原位杂交分析

原产于非洲的埃塞俄比亚芥(Brassica carinata,2n=34,BBCC)具有适应于炎热干旱地区种植等特点,是改良我国芥菜型油菜(B.juncea,2n=36,AABB)的重要种质资源。本研究用基因组原位杂交方法(GISH,Genomic in situ hybridization)分析了芥菜型油菜与埃塞俄比亚芥种间杂种花粉母细胞的染色体分离,发现在后期I染色体主要以17∶18类型分离,其次是16∶19,染色体落后现象偶然发生,其中B染色体组以8∶8的分离比率较高,表明不同来源的B染色体可正常配对分离。本研究表明,芥菜型油菜与埃塞俄比亚芥远缘杂交,通过染色体同源重组(B染色体间),以及部分同源染色体配对交换的方式(A、B、C基因组间),有可能将埃塞俄比亚芥优良遗传成分转移到芥菜型油菜中。     

壳寡糖对干旱胁迫下油菜叶片生理指标的影响

以世界主要油料作物油菜(Brassica napus L.)为研究对象,研究了干旱胁迫下外施壳寡糖对叶片超氧化物歧化酶(SOD)和过氧化物酶(POD)活性、叶绿素荧光参数及叶片相对含水量的影响.结果表明:壳寡糖处理早期油菜叶片SOD和POD活性明显下降,后期SOD和POD活性明显升高;壳寡糖处理后脯氨酸含量明显升高.壳寡糖还能够诱导光系统Ⅱ最大光化学效率(Fv/Fm)、光系统Ⅱ有效光化学量子产量(Fv'/Fm')、光化学荧光猝灭系数(qP)和光系统Ⅱ电子传递量子产量(ΦPsⅡ)明显提高,同时初始荧光(Fo)和非光化学荧光猝灭系数(qN)明显降低.此外,壳寡糖还能够提高油菜叶片相对含水量.研究表明,外施壳寡糖改善了油菜抗旱相关的生理指标进而增强其抗旱能力.     

三峡库区消落带4种典型草本植物的生态化学计量特征

为研究三峡水库消落带优势植物的养分利用特征及其对生境的适应策略,选择消落带分布最多的4种草本植物为研究对象,分析了植物根、茎、叶的碳(C)、氮(N)、磷(P)、钾(K)含量和化学计量比特征。结果表明:(1)相比陆地系统和自然湿地系统,消落带植物具有较低的C含量和较高的N、P、K含量,C/N、C/P、C/K均较低,表明植物具有低固碳和高养分积累、低养分利用效率和高生长速率的特征;(2)4种植物的养分含量和计量比存在一定差异,其中狗牙根具有较低的N、P、K含量和较高的C/N、C/P、C/K,且变异系数均低于其他3种植物,其低养分需求、高养分利用效率以及较强的内稳性可能是其在库区分布最广的重要机制;(3)4种植物在不同器官的养分分配策略相似,均表现为叶片C含量低于根和茎,而N、P、K含量则显著高于根、茎;同时,与根、茎相比,叶片C/N、C/P、C/K较低,N/P、N/K较高,且在不同生境条件下变异系数较小,表现出相对较高的稳定性;(4)落消带植物的养分含量及计量比从全库区上游至下游的空间变异性较强,其中N、C/N、N/P变异性较大,而C、P、K变异性较小,表明植物N含量受生境变化的影响较大,加之消落带不同植物生长均受到严格的N限制,因此N供给可能是影响消落带生态系统结构的重要因子。三峡库区消落带植物生态化学计量特征具有明显的变异性和特殊性,是植物群落演替及生态系统功能变化的重要驱动因素。     

甘蓝型油菜(Eru CMS)与甘蓝种间杂种的同工酶和蛋白质分析

经胚抢救获得甘蓝型油菜(Brassica napus L.)(Eru CMS)与甘蓝(Brassica oleracea L.var.capitata L.)种间杂种,前期经过流式细胞仪、柱头染色体数目、花粉活力等分析获得一些真杂种。利用电泳法,对真杂种植株的3种同工酶(SOD、EST、COD)和蛋白质进行详细分析,了解了杂种与亲本的同工酶和蛋白质的特性差异。结果表明,杂种与亲本之间的同工酶和蛋白质存在较明显的差异:杂种的SOD、COD的酶带表现为偏父本甘蓝型;杂种的EST的酶带表现为偏母本油菜型;杂种的蛋白质电泳表现为不仅具有双亲的特征蛋白带,也有其自身特征蛋白带。     

甘蓝型油菜新型细胞质雄性不育类型NRO4270A的鉴定

甘蓝型油菜细胞质雄性不育系(CMS)NRO4270A是从萝卜甘蓝(Raphanus brassica,RRCC)与甘蓝型油菜(Brassica napus,AACC)远缘杂交后代中发现的一种CMS类型,该研究通过育性调查、恢保关系测定、花药发育的显微观察和线粒体DNA的RFLP分析对NRO4270A不育系进行鉴定。结果表明:(1)通过连续5年在冬油菜区和春油菜区的育性调查,发现NRO4270A不能产生花粉,败育彻底,不育性稳定,不受温度、光照等环境条件的影响,不育率和不育度均为100%。(2)将NRO4270A、pol、ogu、kos、hau等CMS分别与pol、ogu恢复系以及‘华双4号’测交,显示NRO4270A与pol、ogu、kos等不育系统的恢保关系明显不同。(3)通过对NRO4270A与保持系NRO4270B花药发育的显微结构观察和比较,表明NRO4270A花粉的败育时期为四分体期至单核花粉晚期,败育特点为:单核小孢子时期,单核花粉粒不能形成外壁,花药绒毡层液泡化,并逐渐膨大增厚,最后绒毡层和小孢子彻底消失,花粉囊变为空腔,花药完全败育。(4)利用18个探针/酶组合(由6个线粒体基因atp1、atp6、atp9、orf222、cox2、cob探针和3种限制性内切酶EcoRⅠ、BamHⅠ和HindⅢ组成)对NRO4270A、pol、ogu、kos、hau等CMS以及NRO4270B和‘华双5号’的线粒体DNA进行RFLP分析,在11个探针/酶组合中,NRO4270A均显示与其他材料明显不同的带型,表明其为新型的不育胞质类型。NRO4270A的发现和利用将有利于克服中国油菜主要CMS类型对环境的敏感性和生产上不育胞质类型单一化等问题。     

甘蓝型油菜雄性不育系160S育性转换与利用

以甘蓝型油菜雄性不育系160S为试验材料,分别在自然栽种和人工控温条件下对其花器官形态变化、花粉育性转换和杂种优势等进行了初步研究,以探讨植物温敏雄性不育的发生机制。结果表明:环境温度对160S的花器官形态及育性转换具有明显的作用,高温可使花瓣变小,雄蕊退化,花粉活力、角粒数与自交有效结角率降低,表现为低温可育、高温不育,育性变化趋势表现为完全可育-半不育-彻底败育。160S恢复源广泛,且具有较好的配合力和杂种优势,为利用两系法生产油菜杂交种提供了一个较好的途径。     

甘蓝型油菜与埃塞俄比亚芥杂交创制有限花序新种质

远缘杂交是作物遗传改良和种质创新的重要途径。本研究通过甘蓝型油菜(2n=38,AACC)与埃塞俄比亚芥(2n=34,BBCC)的种间杂交获得了形态属于两亲本中间型的杂种F_1,且表现埃塞俄比亚芥的有限花序特性。该杂种具有预期的体细胞染色体数目(2n=36,ABCC),GISH分析显示含有8条B染色体组的染色体;在减数分裂终变期的花粉母细胞中出现多种染色体配对构型,在后期Ⅰ染色体分离多以17∶19分离为主,但在后期Ⅰ及后Ⅱ期均出现大量的落后染色体、染色体桥,导致花粉败育。将该杂种与亲本甘蓝型油菜品种中双11连续回交,获得有限花序的后代材料BC_5F_1,其他形态特征与中双11类似,其体细胞染色体数目恢复至2n=38,减数分裂染色体行为也正常;GISH分析虽检测到少数B染色体组信号,但没有整条的染色体。利用46对B基因组特异引物对有限花序杂种及后代材料进行SSR扩增,发现在杂种F_1及部分BC_5F_1植株中都能检测到B基因组特异带,表明有埃塞俄比亚芥B基因组遗传成分的导入。所创建的有限花序新品系为油菜的遗传改良和选育适于机械化作业的新品种提供新材料。     

玉米CMS-C不育系及其保持系线粒体膜通透性的比较分析

为了解玉米CMS-C相关不同育性材料雄穗线粒体膜通透性表现特征, 本文以玉米细胞质雄性不育系C48-2、保持系N48-2和育性恢复F1 （C48-2×18-599白）雄穗为材料, 测定了线粒体膜相关生理指标。结果发现, 从花粉母细胞时期到双核期C48-2雄穗线粒体膜吸光度、膜电位荧光强度、Ca2＋含量和Cyt c/a比值下降, MDA含量上升, 且各指标均在单核期和双核期与N48-2和F1 （C48-2×18-599白）有显著差异。     

温光型核雄不育小麦育性转换的温度敏感期和临界温度研究

多年人工气候箱和田间分期播种试验表明,小麦温光型隐性核雄不育两用系C49S育性温度敏感期较长,从花粉母细胞形成期到成熟花粉期都有一定程度的敏感性,其中最敏感的时期有两个,即花粉母细胞减数分裂期和小孢子中期,当减数分裂期温度与小孢子中期温度均较低时,C49S表现为高度不育,当其温度均较高时,C49S表现为高度可育;C49S高度不育的临界温度是减数分裂期平均最低温度(Tmin₁)、小孢子中期平均温度(T₂)和平均最低温度(Tmin₂),分别为8.5℃、13.5℃和10.5℃;C49S高度可育的临界温度是Tmin₁11.5℃、T₂15.0℃、Tmin₂12.5℃;对温光型核雄不育两用系在小麦杂种优势利用中的应用价值、条件及风险进行了综合评价.     

亚热带六种天然林树种细根养分异质性

以福建省建瓯市万木林自然保护区常绿阔叶林6种优势树种为研究对象,分析其1—5级细根养分特征及其与细根形态特征和叶片养分的关系。结果发现:除沉水樟细根C含量表现为随序级升高而增加外,其余5种树种均没有明显变化规律;6种树种N、P含量基本表现为随序级升高而降低,C/N随序级升高而增加;但N/P随序级升高均没有明显变化规律。单因素方差分析发现:树种对C、N、P含量和C/N、N/P具有极显著影响(P<0.01),序级对6种树种N、P含量和C/N有极显著或显著影响(P<0.01,P<0.05),对C含量和N/P影响并不显著(P>0.05);双因素方差分析发现:树种和序级的交互作用对C、N、P含量和C/N有极显著或显著的影响(P<0.01,P<0.05),对N/P影响不显著(P>0.05)。相关性分析表明:在较高级根中C、N含量之间呈现相关性,在较低级根中N、P含量呈现相关性,同时C/N、N/P的变化均主要由N含量变化决定;6种树种细根比根长和N、P含量、C/N有极显著相关性(P<0.01),而与C含量和N/P相关性并不明显。一级根的N、P含量和N/P相比于整个细根更接近于叶片,但一级根与叶片N、P、N/P之间并没有显著的相关性(P>0.05)。     

Brassicaceous Seed Meals as Soil Amendments to Suppress the Plant-parasitic Nematodes Pratylenchus penetrans and Meloidogyne incognita

Brassicaceous seed meals are the residual materials remaining after the extraction of oil from seeds; these seed meals contain glucosinolates that potentially degrade to nematotoxic compounds upon incorporation into soil. This study compared the nematode-suppressive ability of four seed meals obtained from Brassica juncea 'Pacific Gold', B. napus 'Dwarf Essex' and 'Sunrise', and Sinapis alba 'IdaGold', against mixed stages of Pratylenchus penetrans and Meloidogyne incognita second-stage juveniles (J2). The brassicaceous seed meals were applied to soil in laboratory assays at rates ranging from 0.5 to 10.0% dry w/w with a nonamended control included. Nematode mortality was assessed after 3 days of exposure and calculated as percentage reduction compared to a nonamended control. Across seed meals, M. incognita J2 were more sensitive to the brassicaceous seed meals compared to mixed stages of P. penetrans. Brassica juncea was the most nematode-suppressive seed meal with rates as low as 0.06% resulting in > 90% suppression of both plant-parasitic nematodes. In general B. napus 'Sunrise' was the least nematode-suppressive seed meal. Intermediate were the seed meals of S. alba and B. napus 'Dwarf Essex'; 90% suppression was achieved at 1.0% and 5.0% S. alba and 0.25% and 2.5% B. napus 'Dwarf Essex', for M. incognita and P. penetrans, respectively. For B. juncea, seed meal glucosinolate-degradation products appeared to be responsible for nematode suppression; deactivated seed meal (wetted and heated at 70 °C for 48 hr) did not result in similar P. penetrans suppression compared to active seed meal. Sinapis alba seed meal particle size also played a role in nematode suppression with ground meal resulting in 93% suppression of P. penetrans compared with 37 to 46% suppression by pelletized S. alba seed meal. This study demonstrates that all seed meals are not equally suppressive to nematodes and that care should be taken when selecting a source of brassicaceous seed meal for plant-parasitic nematode management.     

Physical mapping of rDNA loci in Brassica species.

The number of major rDNA loci (the genes coding for 18S-5.8S-26S rRNA) was investigated in the economically important Brassica species and their wild relatives by in situ hybridization of an rDNA probe to metaphase chromosomes and interphase nuclei. The diploid species B. nigra (B genome) has two major pairs of rDNA loci, B. oleracea (C genome) has two major pairs and one minor pair of loci, while B. campestris (A genome) has five pairs of loci. Among the three tetraploid species arising from these three diploid ancestors, B. carinata (BBCC genomes) has four loci, B. juncea (AABB genomes) has five major pairs and one minor pair of loci, and B. napus (AACC genomes) has six pairs of loci, indicating that the number of loci has been reduced during evolution. The complexity of the known rDNA restriction fragment length polymorphism patterns gave little indication of number of rDNA loci. It is probable that chromosome rearrangements have occurred during evolution of the amphidiploid species. The data will be useful for physical mapping of genes relative to rDNA loci, micro- and macro-evolutionary studies and analysis of aneuploids including addition and substitution lines used in Brassica breeding programs.     

芥菜型和甘蓝型油菜及其杂种后代种子硫甙组成差异的研究

用高效液相色谱技术分析了芥菜型油菜（Ｂｒａｓｓｉｃａ ｊｕｎｃｅａ）和甘蓝型油菜（Ｂｒａｓｓｉｃａ ｎａｐｕｓ）种间杂种后代及共亲本的硫甙组成。结果表明,与亲本农艺性状相似的后代,那么其硫甙组成与亲本亦相似。各后代中硫甙组成与亲本相比,虽发生了不同管理的变化,但都有倾向母体的趋势。芥菜型油菜烯丙基硫甙含量,２－羟基－３－丁烯基硫甙含量低,而甘蓝型油菜２－羟基３－丁烯基硫甙含量高,烯丙基硫甙含量最低     

铀尾沙对油菜幼苗生长和生理特征的影响

采用沙培盆栽试验,以铀尾沙所占比例分别为0%(CK)、25%(T1)、50%(T2)、75%(T3)、100%(T4)的培养基质,研究了铀尾沙对芥菜型油菜、甘蓝型油菜和白菜型油菜出苗率、幼苗生物量、叶绿素含量、丙二醛(MDA)含量、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、过氧化物酶(POD)、抗坏血酸过氧化物酶(APX)、谷胱甘肽还原酶(GR)活性和还原型谷胱甘肽(GSH)含量的影响.结果表明:不同比例的铀尾沙对油菜出苗率的影响差异不显著(P＞0.05).3种油菜的生物量均随铀尾沙处理量的增加而呈先增后降的现象,甘蓝型油菜和白菜型油菜在T2时生物量最大,芥菜型油菜在T3时生物量最大.MDA含量则呈先降低后升高,甘蓝型油菜和白菜型油菜在T2时MDA含量最低,芥菜型油菜在T3时的MDA含量最低,并与叶绿素含量呈显著的负相关.随铀尾沙处理量的增加,3种类型的油菜SOD、CAT、POD、APX和GR活性逐渐增加,GSH含量也持续升高.铀尾沙对油菜幼苗生长和抗氧化酶系统产生了一定的诱导作用,而这3种油菜也对铀尾沙也表现出较强的耐受能力.     

Brassica napus DNA markers linked to white rust resistance in Brassica juncea

White rust, caused by Albugo candida, is an economically important disease of Brassica juncea mustard. The most efficient and cost effective way of protecting mustard plants from white rust is through genetic resistance. The development of canola quality B. juncea through interspecific crosses of B. juncea with Brassica napus has lead to the introgression of white rust resistance from B. napus into B. juncea. The objective of this study was to identify DNA markers for white rust resistance, derived from the introgressed B. napus chromosome segment, in a BC(3)F(2) population of condiment B. juncea mustard. This segregating population was phenotyped for white rust reaction and used to screen for AFLP markers associated with white rust resistance using bulked segregant analysis. Segregation data indicated that a single dominant gene controlled resistance to white rust. Eight AFLP markers linked to white rust resistance were identified, all derived from B. napus. The B. napus chromosome segment, carrying the white rust resistance gene ( Ac2V(1)), appeared to have recombined with the B. juncea DNA since recombinant individuals were identified. Comparative mapping of the eight B. napus-derived AFLP markers in a typical B. napus mapping population was inconclusive; therefore, the size of the introgressed B. napus fragment could not be determined.     

Tracing B-genome chromatin in Brassica napus x B. juncea interspecific progeny.

We used polymerase chain reaction (PCR) and fluorescence in situ hybridization (FISH) techniques to demonstrate the presence of Brassica B-genome chromosomes and putative B-genome introgressions in B. napus x B. juncea interspecific progeny. The B-genome--specific repeat sequence pBNBH35 was used to generate PCR products and FISH probes. The highest frequencies of viable progeny were obtained when B. napus was the maternal parent of the interspecific hybrid and the first backcross. B-genome--positive PCR assays were found in 34/51 fertile F2 progeny (67%), which was more than double the proportion found in fertile BC(1) progeny. Four B-genome--positive F(2)-derived families and 1 BC(1)-derived family were fixed or segregating for B. juncea morphology in the F(4) and BC(1)S(2), respectively, but in only 2 of these families did B. juncea-type plants exhibit B. juncea chromosome count (2n = 36) and typical B-genome FISH signals on 16 chromosomes. The remaining B. juncea-type plants had B. napus chromosome count (2n = 38) and no B-genome FISH signals, except for 1 exceptional F(4)-derived line that exhibited isolated and weak B-genome FISH signals on 11 chromosomes and typical A-genome FISH signals. B. juncea morphology was associated with B-genome--positive PCR signals but not necessarily with 16 intact B-genome chromosomes as detected by FISH. B-genome chromosomes tend to be eliminated during selfing or backcrossing after crossing B. juncea with B. napus, and selection of lines containing B-genome chromatin during early generations would be promoted by use of this B-genome repetitive marker.     

Analysis of B-genome chromosome introgression in interspecific hybrids of Brassica napus × B. carinata

Brassica carinata, an allotetraploid with B and C genomes, has a number of traits that would be valuable to introgress into B. napus. Interspecific hybrids were created between B. carinata (BBCC) and B. napus (AACC), using an advanced backcross approach to identify and introgress traits of agronomic interest from the B. carinata genome and to study the genetic changes that occur during the introgression process. We mapped the B and C genomes of B. carinata with SSR markers and observed their introgression into B. napus through a number of backcross generations, focusing on a BC(3) and BC(3)S(1) sibling family. There was close colinearity between the C genomes of B. carinata and B. napus and we provide evidence that B. carinata C chromosomes pair and recombine normally with those of B. napus, suggesting that similar to other Brassica allotetraploids no major chromosomal rearrangements have taken place since the formation of B. carinata. There was no evidence of introgression of the B chromosomes into the A or C chromosomes of B. napus; instead they were inherited as whole linkage groups with the occasional loss of terminal segments and several of the B-genome chromosomes were retained across generations. Several BC(3)S(1) families were analyzed using SSR markers, genomic in situ hybridization (GISH) assays, and chromosome counts to study the inheritance of the B-genome chromosome(s) and their association with morphological traits. Our work provides an analysis of the behavior of chromosomes in an interspecific cross and reinforces the challenges of introgressing novel traits into crop plants.     

High-oleic acid Australian Brassica napus and B. juncea varieties produced by co-suppression of endogenous Delta12-desaturases

Genetic engineering methods have been used successfully to modify the fatty acid profile of elite Australian germplasm of Brassica napus and B. juncea. Co-suppression plasmids carrying oleate desaturase genes from each species have been constructed and transferred into Australian elite breeding lines of B. napus and B. juncea using Agrobacterium tumifaciens plant-transformation techniques. Modifications to existing Brassica transformation protocols and the use of an intron-interrupted hygromycin-resistance gene as the selectable marker have resulted in improved transformation efficiencies. Silencing of the endogenous oleate desaturase genes has resulted in substantial increases in oleic acid levels, up to 89% in B. napus and 73% in B. juncea.     

Karyotyping of Brassica amphidiploids using 5S and 25S rDNA as chromosome markers

Species of Brassica have small, morphologically similar chromosomes, which makes karyotyping difficult using conventional cytogenetic methods. Molecular cytogenetic methods, like fluorescence in situ hybridisation (FISH) have the potential to improve karyotyping through the use of chromosome- or genome-specific markers. Simultaneous application of more than one DNA probe can greatly enrich the results obtained compared with separate single target FISH experiments. This paper demonstrates the use of multicolour fluorescence in situ hybridisation with 5S and 25S rDNA for karyotyping three amphidiploid species: B. napus, B. juncea and B. carinata. Using this method, it was possible to identify eight out of nineteen pairs of chromosomes in B. napus, ten out of eighteen pairs in B. juncea and six out of sixteen pairs in B. carinata. Additionally, rDNA sites allow the determination of the genomic origin of all marked chromosomes in B. napus and B. juncea.     

Cloning of dehydrin coding sequences from Brassica juncea and Brassica napus and their low temperature-inducible expression in germinating seeds.

A novel subclass of dehydrin genes, homologous to the Raphanus sativus late embryogenesis-abundant (LEA) protein (RsLEA2) and the Arabidopsis thaliana dehydrin, was isolated from Brassica juncea and Brassica napus, here designated BjDHN1 and BnDHN1, respectively. The cDNA of BjDHN1 and BnDHN1 genes share 100% nucleotide identity. The encoded protein is predicted to consist of 183 amino acid residues (molecular mass of 19.2 kDa and pI of 7.0). It shares 85.3% and 65.4% amino acid sequence identity with the RsLEA2 and Arabidopsis dehydrin, respectively. This Brassica dehydrin also features a "Y(3)SK(2)" plant dehydrin structure. Expression analysis indicated that the Brassica dehydrin gene is expressed at the late stages of developing siliques, suggesting that the gene expression may be inducible by water-deficit. Analysis of gene expression also indicated that in germinating seeds the gene expression was inducible by low temperature. Seed germination under low temperature was compared between B. juncea and B. napus. The results showed that B. juncea seeds germinated faster than B. napus seeds. Expression of Brassica dehydrin gene was also examined as a function of seed germination under low temperature.