能源植物小桐子赤霉素合成代谢及信号转导相关基因的鉴定及序列分析

赤霉素(gibberellin,GA)是一类非常重要的植物激素,在植物种子萌发、茎干伸长、叶片生长、腺毛发育、花粉成熟、开花诱导和果实成熟等生长发育过程中都发挥着重要的作用。GA在一年生草本植物中可以促进开花,而在大多数多年生木本植物中则抑制成花诱导。为了更好地研究赤霉素在木本油料能源植物小桐子(Jatropha curcas)开花调控方面的作用机理,我们对小桐子整个基因组中参与GA合成代谢和信号转导的全部基因进行了鉴定和序列分析。这些基因包括6个多基因家族编码的蛋白,即GA2氧化酶(GA2-oxidase,GA2ox)、GA3氧化酶(GA3-oxidase,GA3ox)、GA20氧化酶(GA20-oxidase,GA20ox)、GID1(GIBBERELLIN INSENSITIVE DWARF1)、DELLAs和F-box蛋白,以及2个单基因编码的蛋白,EL1(EARLY FLOWERING1)和SPY(SPINDLY)。采用拟南芥和水稻中已经鉴定的上述基因编码的蛋白序列在小桐子基因组序列数据库和本实验的小桐子转录组数据库中进行BLASTP分析,找到17个同源蛋白的全长序列,并将其与28个拟南芥的、16个水稻的、24个葡萄的和22个蓖麻的同源蛋白构建系统发育树进行比对分析。结果表明,小桐子中参与赤霉素合成代谢及信号转导的大多数基因与蓖麻和葡萄同源基因的相似度更高。     

赤霉素、钙和甜菜碱对小桐子种子萌发及幼苗抗低温和干旱的影响

小桐子(Jatropha curcas L.)属大戟科( Euphorbiaceae)麻疯树属(Jatropha L.)的能源植物。分别以小桐子种子和幼苗为实验材料,研究了不同浓度的赤霉素和CaCl₂单独处理,以及不同浓度的赤霉素、CaCl₂和甜菜碱组合处理对小桐子种子萌发及幼苗抗低温和干旱的影响。结果表明:分别用10 mg/L赤霉素和10 mmol/L CaCl₂处理小桐子种子,不仅可以提高其在正常萌发条件(26℃)、低温(18℃)和干旱胁迫(5% PEG6000)下的发芽率,还可缓解小桐子幼苗在低温(2℃)或干旱胁迫(25% PEG6000)下电解质渗漏率的增加和丙二醛(MDA)的积累。用10 mg/L赤霉素、5 mmol/L CaCl₂和15 mmol/L甜菜碱组合处理小桐子种子,也可进一步增强其在正常萌发(26℃)、低温(18℃)和干旱胁迫(5% PEG6000)条件下种子的发芽率,以及缓解小桐子幼苗在低温(2℃)或干旱胁迫(25% PEG6000)下电解质渗漏率的增加和MDA的积累,表明10 mg/L赤霉素和10 mmol/L CaCl₂分别处理或10 mg/L赤霉素、5 mmol/L CaCl₂和10 mmol/L甜菜碱组合处理可提高小桐子种子在低温和干旱胁迫下的发芽率,以及提高小桐子幼苗对低温和干旱胁迫的抵抗能力。     

小桐子愈伤组织的诱导

本文以小桐子的叶片、叶柄、茎段及下胚轴和子叶作为外植体,研究不同外植体类型对愈伤组织诱导的影响,结果表明叶柄的诱导率最高,其次为茎段的诱导率。同时以小桐子的下胚轴作为外植体,研究植物生长调节剂种类及浓度配比对愈伤组织诱导的影响,结果显示6-BA与2,4-D的组合更适宜小桐子愈伤组织的诱导,MS+6-BA0.5mg/L+2,4-D0.1mg/L为小桐子愈伤组织诱导的最佳培养基,其愈伤组织诱导率高达96.7%。本研究为小桐子愈伤组织的分化、植株再生及相关的遗传转化研究提供了参考。     

小桐子的组织培养和植株再生

以小桐子（Jatropha curcas）的胚芽、子叶、下胚轴、叶柄、叶片和茎段作为外植体,用不同浓度的6-苄基腺嘌呤（6-BA）和α-萘乙酸（NAA）对其进行愈伤组织的诱导和植株再生的研究。结果表明：在MS培养基中加入5．0mg／L6-BA和1．0mg／LNAA对愈伤组织的诱导效果最好;加入5．0mg／L6-BA和0．1mg／LNAA对不定芽的诱导最为有效,加入0．1mg／L6-BA和1．0mg／LNAA有利于芽的生长;加入1．0mg／LNAA的1／2MS培养基对生根最为有利。     

利用小桐子饼生产有机肥是小桐子饼利用的重要途径

饼粕作为小桐子生物柴油生产过程中产生的最大宗副产物,能否有效利用对小桐子生物柴油产业的发展具有极其重要的作用.小桐子饼含有丰富的营养物质,将其发酵制成有机肥是人们最常采用的方法.通过对小桐子饼发酵生产有机肥的现状、技术路线和经济价值进行深入分析,认为小桐子饼发酵生产有机肥是一条环保、经济适用的利用小桐子饼的技术路线,对...     

小桐子花的性别与开花时间的相互关系

通过对小桐子(Jatropha curcas L.)花序上小花的开花顺序和小花性别的观察,发现其聚伞花序中心花的性别与其开花时间相关联,同时还和周围其它小花的性别有很大的相关性.中心花若在整个花序开花的第一天开放,仅有23.2%的可能是雌花;若在第二天及以后开放,有85%以上的可能为雌花.而中心花的性别又与其它花的性别相关,中心花如为雌花,那么聚伞花序上的其它小花有80%的可能性为雌花;但中心花若为雄花,那么整个聚伞花序上将不会再有雌花.研究小桐子的开花行为及性别控制规律,对于进一步人工调控小桐子花的性别表达,提高小桐子的产量、培育高产稳产品种,具有重要的应用价值.     

云南南部不同种源地小桐子遗传多样性的ISSR分析

应用ISSR分子标记方法对采自云南的8个居群的小桐子(Jatropha curcas)共158个个体进行遗传多样性分析。8个ISSR引物共扩增到了67个位点,其中61个是多态性位点。分析结果表明:(1)云南小桐子的遗传多样性水平很高。在物种水平上,平均每个位点的多态位点百分率PPB=91.04%,有效等位基因数Ne=1.5244,Nei′s基因多样性指数He=0.3070,Shannon多样性信息指数Ho=0.4618;在居群水平上,PPB=55.04%,Ne=1.3826,He=0.2171,Shannon多样性信息指数Ho=0.3178。(2)居群间的遗传分化低于居群内的遗传分化。基于Nei′s遗传多样性分析得出的居群间遗传多样性分化系数Gst=0.2944。AMOVA分析显示:云南小桐子的遗传变异主要存在于居群内,占总变异的63.50%,居群间的遗传变异占36.50%。(3)居群间的地理距离及遗传一致度并不存在相关性。鉴于以上指标,我们推测云南小桐子可能来自不同的地区。     

小桐子内生真菌及其抗真菌活性研究

从药用植物小桐子的根和茎中分离到内生真菌57株,经鉴定,它们分属于15个不同的分类单元。其中拟盘多毛孢为优势属,其次为拟茎点霉属和茎点霉属。用杨桃炭疽菌,香蕉疫霉和镰刀菌作指示菌对这些内生真菌的抗真菌活性进行检测,结果表明,有2株菌对杨桃炭疽菌具有抑菌活性。     

土施多效唑对小桐子营养生长与生殖生长的影响

小桐子(Jatropha curcas)是一种具有多种用途的植物,其种子含油量高达40％,是一种很有应用前景的能源植物.小桐子在温度、湿度较高的地区,特别是其修剪后新萌发的枝条经常出现营养生长旺盛,成花量较少的现象,导致其种子产量较低.本研究采用多效唑(paclobutrazol,PAC)对小桐子进行土施处理,结果表明:多效唑的使用剂量为0.8g·m-1树冠直径时,可有效抑制小桐子的营养生长,降低株高;同时促进小桐子的生殖生长,增加开花和结果枝条数量,增加花序和果序数量,增加单个花序的总花数和雌花的比例,可将小桐子单株种子产量提高2.4倍.此外,多效唑处理使小桐子的开花期和结果期更加集中,便于田间管理和采收.多效唑作为赤霉素合成的抑制剂,可以有效地促进小桐子由营养生长向生殖生长的转变,对于了解赤霉素在调控小桐子花发育方面的作用具有重要意义;同时也有助于克隆有关功能基因,进一步采用转基因技术对小桐子进行遗传改良.     

云南小桐子资源调查与评价

对云南省小桐子（Jatropha curcas）资源分布、环境因子和种子相关性状展开了调查和分析,结果表明：所测居群的种子平均千粒重、出仁率、种仁含油率和种子含油率分别为567．9克、56．67％、61．78％和35．13％。居群间种子千粒重、出仁率、种仁含油率、种子含油率都有极显著的差异,丽江A、红河D、丽江C、丽江B和玉溪A等4个居群有较高开发利用价值。温度是影响云南省小桐子地理分布的主要限制因子,分布区的海拔高度和年均温有强负相关性。云南省最适合种植小桐子的地区是金沙江和元江流域的干热河谷地区,其次是临沧、保山、普洱、西双版纳等地的高温低海拔地区。本研究对云南省小桐子的研究和产业发展提出了建议。     

The incidence and severity of Jatropha dieback disease in Zaria,Nigeria

A survey was conducted on Jatropha plantations within Zaria to assess disease incidence and severity on two fields along Shika road and near the dam (Irrigation site) of the Institute for Agricultural Research (IAR) at Samaru and a plantation of the National Research Institute for Chemical Technology (NARICT) at Basawa. An assessment of incidence and severity of the disease was done on six accessions collected from four Local Government Areas (Giwa, B/Gwari, Soba, Lere) of Kaduna state and grown in one of the I.A.R fields along Bomo road at Samaru. The accessions were collected from Gangara, Maidaro, Farin Gida (Giwa LGA), Birnin Gwari (B/Gwari LGA), Kayarda (Lere LGA) and Soba (Soba LGA). The causal organism of dieback on Jaropha curcas L, was isolated and its pathogenicity determined. The isolated Fusarium sp. incited dieback/stem-rot disease when inoculated on healthy Jatropha seedlings. The disease incidence was significantly higher on NARICT plantation compared to the two IAR fields which were not significantly different from each other. The accessions from Gangara, Kayarda, Birnin Gwari and Soba did not significantly differ from each other but had significantly higher disease incidence than those from Maidaro and Farin Gida. Accession from Soba had significantly higher disease severity compared to the accessions from all the other LGAs, which did not differ statistically from each other.     

Disease progress of Jatropha dieback caused by Fusarium sp. in Samaru,Zaria

A survey was conducted from June 2012 to May 2013 in the Institute for Agricultural Research Jatropha plantation along Shika road to determine the progress of incidence and severity of Jatropha dieback caused by Fusarium sp. over a year. The disease incidence did not significantly differ among the months under review while disease severity was highest in August and lowest in March 2013.     

麻疯树中ADP核糖基化因子基因的克隆和表达

从麻疯树cDNA文库中筛选到包含完整编码序列的ADP核糖基化因子的cDNA序列,命名为Jc-arf．其长度为887bp,开放阅读框546bp,推测的编码蛋白含181个氨基酸残基,具有典型的GTP结合蛋白家族的特点：P框（GLDAAGKT）、G’框（DVGGQ）和G框（NKQDL）。序列分析显示,Jc-arf矿接近于人类ClassI型arf基因,其蛋白序列与多种植物ARF有很高的同源性。半定量RT-PCR结果显示,Jc-arf的表达具有一定的组织特异性,在花中最丰富。PEG、低温、NaCl胁迫下,Jc-arf的表达受PEG和低温的诱导,而对NaCl胁迫不敏感。     

用GC-MS分析不同采收和贮存时期的麻疯树种子油的脂肪酸

将采收于青果期、黄果期、黑果期及储存1a、2a的麻疯树种子提油,测定其理化性质,并利用GC-MS分析这5个不同时期的种子所提取的麻疯树种子油成分。结果表明:5个油样的水分、酸值、出油率有较大差异,主要化学成分、碘值、皂化值差异不大。青果的出油率为13.13%,水分为0.66%,酸值为69.21,不饱和脂肪酸含量最低。新采成熟果实的出油率为54.64%,水分为0.36%,酸值为1.51,不饱和脂肪酸含量也相对较高。因此,新采的成熟果实较为适合作生物柴油的原料。     

The expression of the salt-responsive gene salT from rice is regulated by hormonal and developmental cues

The expression pattern of the salT gene was analyzed in different cell types and organs of rice (Oryza sativa L.) in response to saline and hormonal treatments to obtain detailed information on the physiological cues controlling gene expression. Gel blot analysis of RNA and in-situ hybridization performed on seedlings grown for 10 ds in the presence of 1% NaCl revealed that salT was expressed mainly in the younger tissues of the plant. In contrast, 6-week-old plants exhibited maximal salT mRNA accumulation in sheaths of older leaves. In addition, salT was normally expressed in rapidly dividing suspension-cultured cells, but not in quiescent ones. Altogether, these results may indicate that salT expression in each region of the plant is dependent on the metabolic activity of the cells as well as on whether or not they are stressed. The effects of two growth regulators, abscisic acid (ABA) and gibberellic acid, were investigated in combination with the effects of NaCl. Gibberellic acid had a synergistic effect on the induction of the salT gene when combined with 0.5% NaCl, but did not induce salT on its own. At 10 μM, ABA induced salT both in the absence of NaCl and in its presence. Whereas 1 μM ABA acted additively with NaCl to induce gene expression, 5 μM ABA with NaCl was only as effective as NaCl alone. This may indicate that the two stimuli act independently and possibly through antagonistic signal transduction pathways. Received: 26 March 1998 / Accepted: 11 July 1998     

膏桐种子形态、粗脂肪含量与环境因子关系

对云南省两个不同地区膏桐（Jatropha curcas）种子形态特征,粗脂肪含量与环境因子关系开展研究,结果表明：湿热环境的景洪较干热环境的元谋膏桐种子成熟散布期提前,形态特征也优于元谋膏桐种子（P〈0．01）;土壤有机质是影响膏桐种子粗脂肪积累最重要的因子,对膏桐种子粗脂肪积累起显著正效应;干旱胁迫也是影响膏桐种子粗脂肪积累的一个重要的限制因子;景洪膏桐种子粗脂肪含量、含水量较元谋膏桐种子更高,但粗脂肪含量与含水量比率较低（P〈0．01）;具有较高粗脂肪含量的膏桐种子与其含水量的比率可能是植物对干旱胁迫适应的一种机制,其对物种的适应有着重要的生态学意义。     

Analysis of expressed sequence tags from biodiesel plant Jatropha curcas embryos at different developmental stages

Jatropha curcas is considered a potential biodiesel feedstock plant whose seeds contain up to 40% oil. However, little is currently known about the seed biology of Jatropha. Therefore, it would be valuable to understand the mechanisms of development and lipid metabolism in Jatropha seeds. In the present study, three cDNA libraries were constructed with mRNA from Jatropha embryos at different stages of seed development. A total of 9844 expressed sequence tags (ESTs) were produced from these libraries, from which 1070 contigs and 3595 singletons were obtained. One hundred and seven unigenes were found to be differentially expressed in the three cDNA libraries of Jatropha embryos, indicating that these genes may play key roles in seed development. We have identified 59 and 61 unigenes that might be involved in the development and lipid metabolism in Jatropha seeds, respectively. Some of these genes may also play important roles in embryogenesis, morphogenesis, defense response and adaptive mechanisms in plants.