小麦春化发育的分子调控机理研究进展

春化发育特性是小麦品种的重要性状,直接影响着小麦品种的种植范围和利用效率.本文就小麦春化相关基因的发现,以及对春化相关基因VRN1、VRN2和VRN3的克隆、表达特性以及春化发育分子调控机理方面的研究进展进行了综述.     

植物春化特性及春化作用机理

春化作用是某些高等植物成花转变的重要环节,被认为是植物在低温诱导下促使其相关基因的表达,从而导致生理状态转变的一种受遗传控制的生理过程.本文对植物春化反应特性、春化作用的生理生化特性及春化作用的分子生物学等方面的研究进展进行了概述,并对春化研究中的问题进行了分析和展望.     

高羊茅春化基因FaVRN1亚细胞定位与差异表达分析

高羊茅在生长季出现生殖枝,抑制新枝形成,不利于草坪质量及其持久性生长.研究春化基因的分子特征,探索抑制生殖生长的分子育种新途径,对坪用型高羊茅品种改良具有重要意义.本研究在克隆高羊茅春化基因FaVRN1的基础上,构建高羊茅春化基因FaVRN1与绿色荧光蛋白基因GFP融合的植物表达载体p-FaVRN1-hGFP,利用基因枪转化法转入洋葱表皮细胞,荧光显微镜检测融合基因的瞬时表达,并运用实时荧光定量PCR分析春化基因FaVRN1在春化与非春化条件下的表达差异.研究结果表明,FaVRN1基因编码的蛋白产物位于细胞核,符合它作为转录因子特性;春化条件下,FaVRN1基因的表达随处理时间延长逐渐增加.非春化条件下,FaVRN1基因的表达随处理时间延长而降低.FaVRN1基因在春化条件下的表达水平远高于非春化条件,FaVRN1基因的表达受春化条件正调控.     

冬小麦春化相关基因cDNA(verc203)片段序列的分析

春化作用是高等植物发育的重要环节之一,它是受遗传控制的生理过程,人们对此现象已作了生态、生理和生化方面的研究（谭克辉1983）。认为植物茎尖生长点接受低温诱导后,会引起体内许多代谢途径和方式的顺序改变（种康和谭克辉1993）,可能导致春化相关基因启动表达。有人（Burn等1993）提出春化相关基因启动表达可能是基因脱甲基化结果的观点。但这一假说尚缺乏直接的强有力证据。违斌和谭克辉（199）系统地研究了冬小麦春化过程中核酸、蛋白质代谢与春化作用的直接相关关系和春化特异蛋白质代谢与特异性rnRNA出现的重要意义,确信春化…     

植物春化特性及春化作用机理

春化作用是某些高等植物成花转变的重要环节,被认为是植物在低温诱导下促使其相关基因的表达,从而导致生理状态转变的一种受遗传控制的生理过程。本文对植物春化反应特性、春化作用的生理生化特性及春化作用的分子生物学等方面的研究进展进行了概述,并对春化研究中的问题进行了分析和展望。     

冬小麦春化作用相关cDNA克隆Vrc79的分子克隆

在低温需求型植物的发育过程中,春化作用是诱导植物成花的必要因子。在冬小麦春化进程中存在着一个核酸代谢的关键期,为了探讨春化作用的分子机理,以不春化及脱春化的冬小麦京冬1号（Ｔｒｉｔｉｃｕｍ　ａｅｓｔｉｖｕｍＬ．ｃｖ．Ｊｉｎｄｏｎｇ　Ｎｏ．１）胚芽的ｍＲＮＡ为对照,通过减法杂交构建了富集冬小麦春化相关基因的ｃＤＮＡ文库,经过差异筛选分离到了一个仅在春化２１ｄ这一关键期表达,而在不春化、春化４ｄ、脱春化不表达的春化相关。ｃＤＮＡ克隆Ｖｒｃ７９。Ｎｏｒｔｈｅｍ杂交及同源性分析表明它是一个在植物中新发现的不同于低温胁迫诱导基因的春化特异克隆,其对春化需求型植物的成花诱导可能起重要的调控作用。     

拟南芥春化作用相关基因FLC的表达调控及天然早花突变体的研究进展

植物开花是从营养生长到生殖状态的重要发育转变,是多种内在因子和环境因素共同作用的结果。在拟南芥开花调控网络中,开花抑制基因FLC处于枢纽地位。FLC的表达受许多来自环境和生长发育的信号调控,主要包括：PAF1复合体、SWR1复合体成员,FRI依赖途径、自主途径和春化作用途径基因。本文主要综述了影响FLC表达的春化相关基因及天然早花突变体的研究进展,并根据最新的研究成果提出该研究领域的研究方向和重点。     

植物非编码RNA调控春化作用的表观遗传

在自然界中许多高等植物需要通过冬季的低温阶段实现从营养生长到生殖生长的时期转化,这一生物学过程称作春化作用。小麦(Triticum aestivum L.)和油菜(Brassica napus L.)等作物以种子为产品器官,生产上往往通过茬口安排和栽培措施使植株尽早通过春化作用,以促进花芽形成和花器官发育,而大白菜(B rapa ssp.pekinenesis)和甘蓝(B.oleracea)等作物以叶球等营养器官作为产品器官,生产上则设法避免低温引起的春化作用,以保证产品器官的充分生长。FLOWERING LOCUS C(FLC)作为一种重要的开花抑制蛋白负调控春化作用,参与植株从营养生长向生殖生长的转化过程。文章综述了春化中FLC表达受抑制主要通过低温诱导表达FLC基因区域的非编码RNA以及VRN1、VRN2、VIN3等蛋白参与介导组蛋白甲基化,从而在表观遗传上控制春化作用的进程和产品器官的正常发育。     

冬性植物红菜薹在不同温度处理下花青素积累的分子机制(英文)

芸薹属植物红菜薹(Brassica rapa)是一种常见的蔬菜,它的花茎和叶柄表皮中均积累有花青素。为了解红菜薹中花青素合成的分子机制,进行了花青素含量的测定和花青素合成相关基因的表达分析。研究结果表明,叶柄表皮中的花青素含量显著高于叶片(去主脉)的花青素含量。同时,叶柄表皮花青素合成相关基因的表达水平高于叶柄(去表皮)和叶片(去主脉)的表达水平,这表明红菜薹中花青素的合成调控发生在转录水平。BrMYBA1仅在叶柄表皮中表达,但BrbHLH1和BrWD40在叶片和叶柄表皮中均能检测到表达。因此,BrMYBA1的转录激活可能与红菜薹的花青素合成相关。连续低温处理时,红菜薹叶柄表皮中的花青素含量逐渐增加,而该组织中花青素合成的结构基因表达水平逐渐降低。     

冬性植物红菜薹在不同温度处理下花青素积累的分子机制

芸薹属植物红菜薹（Brassica rapa）是一种常见的蔬菜,它的花茎和叶柄表皮中均积累有花青素。为了解红菜薹中花青素合成的分子机制,进行了花青素含量的测定和花青素合成相关基因的表达分析。研究结果表明,叶柄表皮中的花青素含量显著高于叶片（去主脉）的花青素含量。同时,叶柄表皮花青素合成相关基因的表达水平高于叶柄（去表皮）和叶片（去主脉）的表达水平,这表明红菜薹中花青素的合成调控发生在转录水平。BrMYBA1仅在叶柄表皮中表达,但BrbHLH1和BrWD40在叶片和叶柄表皮中均能检测到表达。因此,BrMYBA1的转录激活可能与红菜薹的花青素合成相关。连续低温处理时,红菜薹叶柄表皮中的花青素含量逐渐增加,而该组织中花青素合成的结构基因表达水平逐渐降低。     

芸苔属蔬菜小孢子胚状体再生成苗及倍性鉴定

研究影响大白菜、甘蓝和红菜薹小孢子胚状体再生成苗的几个生理因素的结果表明,在1．0％～1．2％的琼脂中胚状体再生成苗率显著高于0．8％琼脂的。4℃处理10d可显著提高大白菜和甘蓝胚状体再生成苗率。大白菜和红菜薹胚状体再生成苗的最适胚龄为20—29d,甘蓝则为30—35d。培养基B,和MS对再生成苗率影响不大。检测3种芸苔属蔬菜小孢子再生植株的倍性结果表明,大白菜和红菜薹小孢子植株自然加倍率较高,均超过70％;甘蓝较低,仅为30％左右。同一物种的不同品种间胚状体再生成苗所需的条件和加倍效率基本一致。     

Genetic and physical mapping of flowering time loci in canola (Brassica napus L.)

We identified quantitative trait loci (QTL) underlying variation for flowering time in a doubled haploid (DH) population of vernalisation—responsive canola (Brassica napus L.) cultivars Skipton and Ag-Spectrum and aligned them with physical map positions of predicted flowering genes from the Brassica rapa genome. Significant genetic variation in flowering time and response to vernalisation were observed among the DH lines from Skipton/Ag-Spectrum. A molecular linkage map was generated comprising 674 simple sequence repeat, sequence-related amplified polymorphism, sequence characterised amplified region, Diversity Array Technology, and candidate gene based markers loci. QTL analysis indicated that flowering time is a complex trait and is controlled by at least 20 loci, localised on ten different chromosomes. These loci each accounted for between 2.4 and 28.6 % of the total genotypic variation for first flowering and response to vernalisation. However, identification of consistent QTL was found to be dependant upon growing environments. We compared the locations of QTL with the physical positions of predicted flowering time genes located on the sequenced genome of B. rapa. Some QTL associated with flowering time on A02, A03, A07, and C06 may represent homologues of known flowering time genes in Arabidopsis; VERNALISATION INSENSITIVE 3, APETALA1, CAULIFLOWER, FLOWERING LOCUS C, FLOWERING LOCUS T, CURLY LEAF, SHORT VEGETATIVE PHASE, GA3 OXIDASE, and LEAFY. Identification of the chromosomal location and effect of the genes influencing flowering time may hasten the development of canola varieties having an optimal time for flowering in target environments such as for low rainfall areas, via marker-assisted selection.     

The Interaction of Plant Growth Regulators and Vernalization on the Growth and Flowering of Cauliflower (Brassica oleracea var. botrytis)

The growth and flowering response of a cold-requiring cauliflower (Brassica oleracea var. botrytis cv. 60 day) to a range of temperatures under 10 h photoperiod and to growth regulator application were investigated. Endogenous gibberellin A₁(GA₁) concentrations were also assessed under these treatments. Flowering and growth of the inflorescence stalk were correlated with plant developmental stage at the time of a vernalizing cold treatment. Temperature and its duration also affected flowering and inflorescence development. The most effective temperature for inflorescence induction was 10 °C. Flowering did not occur in non-vernalized plants (25 °C) even though they had been treated with GA₃. Application of GA₃ promoted inflorescence stalk elongation greatly in vernalized plants (10 °C), but less so in partially vernalized plants (15 °C or 20 °C). Paclobutrazol (PP₃₃₃) sprayed at the 8–9 leaf stage significantly suppressed inflorescence stalk length and slightly delayed flower bud formation and anthesis. Vernalization at 10 °C increased endogenous GA₁ content in both leaves and the inflorescence stalk irrespective of GA₃ or PP₃₃₃ treatment. Application of GA₃ tended to increase GA₁ levels, while PP₃₃₃ significantly reduce GA₁, both irrespective of vernalization. Vernalization is an important factor for flowering, but not curd formation in this cauliflower cv. 60 day and GA₁ is likely a causal factor in inflorescence stalk elongation.     

Agrobacterium tumefaciens-mediated transformation of broccoli (Brassica oleracea var. italica) and cabbage (B. oleracea var. capitata)

Transgenic broccoli (Brassica oleracea var. italica) was produced by two Agrobacterium tumefaciens-mediated transformation methods. One used flowering stalk explants from mature plants; the other used hypocotyl and petiole explants from in vitro-grown seedlings. Several hundred transformants containing a Bacillus thuringiensis -endotoxin gene (CryIA(c)-type) and the neomycin phosphotransferase gene were recovered. Rooted transformants were obtained in as little as 3 months using seedling explants. Transgenic cabbage was also obtained by the seedling explant method. Parameters important for high efficiency regeneration and transformation rates included use of a tobacco nurse cell layer, sealing of petri dishes with a porous surgical tape instead of Parafilm, preculture of seedling explants and appropriate length of co-cultivation with Agrobacterium. Advantages and disadvantages of each transformation procedure are discussed.     

菜心BrcuFCA基因的克隆与表达分析

本文以菜心(Brassica campestris L.ssp.chinensis(L.)Makino var.utilis)为实验材料,比较了Trizol法、改进Trizol法及改良SDS法提取菜心总RNA的效果。结果表明改进Trizol法集中了操作简单、耗时短、所需试剂种类少,且完整性好,产量和纯度均高等多重优点,成为提取菜心总RNA首选方案。同时文章以甘蓝型油菜及其它物种的FCA保守区设计引物,将质量高的菜心总RNA反转录合成cDNA,通过RT-PCR从菜心的早、晚熟品种中均克隆得到与开花有关的基因片段,将之命名为BrcuFCA,片段长1001bp,GenBank登录号为EU700363,与甘蓝型油菜及拟南芥FCA氨基酸同源性分别达到了98%和82%。另一方面,本研究还利用半定量RT-PCR研究了BrcuFCA不同时空表达特性,并结合已发表的开花调控遗传途径中两个关键基因BrcuFLC和BrcuFRI时空表达特性研究结果,推测菜心主要的抽薹开花途径不是春化和FRI-依赖途径,而很可能是自主开花途径。本研究为菜心抽薹开花分子机理的研究提供理论依据。     

Comparison of Flowering Time Genes in Brassica Rapa, B. Napus and Arabidopsis Thaliana

The major difference between annual and biennial cultivars of oilseed Brassica napus and B. rapa is conferred by genes controlling vernalization-responsive flowering time. These genes were compared between the species by aligning the map positions of flowering time quantitative trait loci (QTLs) detected in a segregating population of each species. The results suggest that two major QTLs identified in B. rapa correspond to two major QTLs identified in B. napus. Since B. rapa is one of the hypothesized diploid parents of the amphidiploid B. napus, the vernalization requirement of B. napus probably originated from B. rapa. Brassica genes also were compared to flowering time genes in Arabidopsis thaliana by mapping RFLP loci with the same probes in both B. napus and Arabidopsis. The region containing one pair of Brassica QTLs was collinear with the top of chromosome 5 in A. thaliana where flowering time genes FLC, FY and CO are located. The region containing the second pair of QTLs showed fractured collinearity with several regions of the Arabidopsis genome, including the top of chromosome 4 where FRI is located. Thus, these Brassica genes may correspond to two genes (FLC and FRI) that regulate flowering time in the latest flowering ecotypes of Arabidopsis.     

基于非变性聚丙烯酰胺凝胶电泳的菜心SCoT分析

以菜心(Brassica rapa var. parachinensis)抗小菜蛾品种Caixin65和感小菜蛾品种Caixin69及6份F2株系为材料,利用非变性聚丙烯酰胺凝胶电泳检测菜心抗虫株系的SCoT多态性,同时进行SCoT多态性的非变性聚丙烯酰胺凝胶电泳检测效率、遗传多样性分析和差异条带克隆分析。结果表明,非变性聚丙烯酰胺凝胶能检测出亲本与子代间的SCoT多态性和遗传多样性变化,条带数量较多且清晰,提高了SCoT标记检测效率。选取亲本的10条非变性聚丙烯酰胺凝胶差异片段克隆测序,获得感虫序列4条、抗虫序列6条,GENSCAN预测其中8条具有启动子、终止子、阅读框等基因结构序列。同源性检索分析表明,感虫序列分别与泛素羧基末端水解酶mRNA序列、大白菜克隆序列、白菜线粒体丙酮酸载体蛋白序列、甘蓝基因组编码未知蛋白的HDEM序列同源,抗虫序列分别与白菜RNA假尿苷合酶4线粒体mRNA序列、京水菜线粒体DNA序列、白菜未知蛋白mRNA序列、白菜4-香豆酸：辅酶A连接酶mRNA序列、大白菜克隆序列、哈茨木霉mRNA序列同源。本研究提高了SCoT标记清晰度、遗传多样性检测水平和差异片段克隆的精准性,使得SCoT成为批量克隆差异片段的高效工具,有助于挖掘SCoT功能性标记信息,开展初步的功能基因组学研究,提高优异株系筛选鉴定效率,加快育种进程,为菜心抗虫性机制的进一步研究提供理论基础。     

硅对菜心炭疽病发生、菜薹形成及硅吸收沉积的影响

采用不同浓度硅(Si)对菜心进行处理, 研究了Si对菜心炭疽病发生、菜薹形成及Si吸收沉积的影响.结果表明：Si对菜心炭疽病的发生有明显的调控作用,并存在基因型差异;感病品种2.5 mmol·L^-1和抗病品种0.5 mmol·L^-1Si处理对炭疽病菌的抗性最强,病情指数最低,菜薹产量最高.Si对菜心品质也有明显影响,不同浓度Si处理可促进感病品种的叶绿素、粗纤维、Vc的合成,并诱导可溶性糖的生成,且叶绿素、粗纤维含量随Si处理浓度的提高而增加;抗病品种的叶绿素含量随Si处理浓度的增加明显增加,而Vc含量下降,Si对抗病品种的粗纤维、可溶性糖含量影响不显著;Si对两个品种菜薹的粗蛋白和可溶性蛋白质含量影响均不显著.施Si明显提高了菜心叶片Si的积累,且叶片的Si含量随着Si处理浓度的增加而显著提高;叶片组织沉积的Si粒大小不一,Si在叶片表皮组织呈不均匀分布.表明Si在菜心叶片中的积累可提高植株的抗病能力,但其积累量与植株的抗病能力不呈直线相关关系.     

Effects of Interactions between Low-temperature Treatments, Gibberellin (GA3) and Photoperiod on Flowering and Stem Height of Spring Rape (Brassica napus var. annua)

Exogenous gibberellin A₃(GA₃) reduced the number of leaf nodesat flowering and time to flowering and increased the stem heightat flowering in three genotypes of spring rape (Brassica napusvar.annua L.). The responses to GA₃were similar to those forlong days (LD) and low-temperature treatments, suggesting thatthe effect of photoperiod and the vernalization response areprobably mediated through gibberellins. The response to exogenousGA₃was greatest in non-cold-treated plants in short days (SD)suggesting that endogenous GAs are limiting in these conditions.CCC, an inhibitor of gibberellin biosynthesis, caused a smallincrease in the number of leaf nodes at flowering and time toflowering and a small decrease in the stem height at flowering,but unexpectedly, its effect was hardly influenced by the applicationof exogenous GA₃. Genotypes that showed the clearest responsesto the treatments with regard to the number of leaf nodes atflowering and time to flowering did not show the clearest responseswith regard to the stem height at flowering; the pattern ofresponses of the number of leaf nodes at flowering and timeto flowering was distinct from that of stem height at flowering.This indicates that flower formation and stem elongation areseparable developmental processes which may be controlled bydifferent endogenous gibberellins, different levels of a specificendogenous gibberellin, or different responses to gibberellin.Copyright 1999 Annals of Botany Company Brassica napus var. annua, gibberellin, photoperiod, spring rape, vernalization.     

Residue dynamics and risk assessment of dimethoate in sweet potato,purple flowering stalk,Chinese kale,celery, and soil

Residue dynamics and risk assessment of the insecticide dimethoate applied to sweet potato, purple flowering stalk, Chinese kale, celery were investigated under the climatic conditions of China. The dissipation experiments indicated that the half-lives of dimethoate in purple flowering stalk, Chinese kale, celery, and soil were 5.9–6.5, 3.8–5.1, 3.5–5.4, 3.4–3.6 d, respectively. The terminal residues of dimethoate and omethoate in the vegetables and soil ranged from 0.008 to 1.73 mg kg^?1 at preharvest intervals of 3, 5, and 7 d. The results showed risk quotient (RQ) of <1 for sweet potato, Chinese kale, and celery, and of >1 for purple flowering stalk when under the age of 18, indicating that spraying dimethoate on sweet potato, Chinese kale, and celery at the recommended dosage is safe for human consumption, whereas spraying it on purple flowering stalk is associated with some risks to human health.