麻疯树种子的研究进展

麻疯树（Jatropha curcas L．）为大戟科（Euphorbiaceae）麻疯树属半肉质小乔木或大灌木,具有很强的抗旱、耐贫瘠的特性。麻疯树的根、树皮、叶和种子均可人药。种子中主要含有脂肪类物质、蛋白质和萜类物质,其毒素为麻疯树毒蛋白和种子油。种仁中的含油量约为50％,可作为理想的生物柴油;毒蛋白、种子油及其他种子提取物可作为生物农药。关于麻疯树种子的发育、脱水行为及其调控研究较少。麻疯树是二种具有重要经济价值的战略资源。     

麻疯树核糖体失活蛋白Curcin2的原核可溶性表达及其抗真菌活性研究

Curcin2是麻疯树幼苗在真菌侵染、干旱及高低温胁迫下诱导产生的一种核糖体失活蛋白.采用分子克隆的方法,从麻疯树基因组中扩增到Curcin2成熟肽编码基因,分别连接到质粒载体pGEX-6p-1、pMAL-c5E上,转化大肠杆菌最终获得重组菌PGC(pGEX-6p-1-curcin2)和PMC(pMAL-c5E-curcin2).在不同温度、IPTG浓度、时间诱导下,Curcin2在重组菌PGC中均以包涵体形式表达,在重组菌PMC中主要以可溶性融合蛋白形式表达,且蛋白质的表达量与诱导条件相关.重组菌PMC表达可溶性curcin2的优化条件为:温度28℃,IPTG 0.3mmol/L,时间8h,此条件下目的蛋白占总蛋白表达量的30.6％,1L培养物中可获得19.74mg电泳纯的重组蛋白.重组蛋白可被MBP TrapTM HP柱亲和纯化并与curcin抗体发生抗原抗体反应.体外抗真菌活性实验表明,纯化后的curcin2融合蛋白有抑制真菌生长作用,且对小麦赤霉、油菜菌核的抑制作用强于curcin.此蛋白的获得为其相关功能的研究奠定了基础.     

麻疯树种油的理化性质及籽粕中元素含量分析及其应用价值初探

麻疯树(Jatropha curcas L. )为大戟科(Euphorbiaceae)麻疯树属(Jatropha L. )植物,果实含油量高,种仁含油量在60%左右,可用于加工提炼优质生物柴油,以替代日益紧缺的石油能源~([1-2]).近年来,由国家科学技术部和联合国开发计划署共同开展的绿色扶贫项目中,麻疯树已经在中国石漠化最为严重的贵州、四川及云南等省区试种成功.麻疯树全株有毒,其种子、树皮、叶、根和乳汁中含有多种化学成分,可作为医药和生物农药资源;其种子加工后残留的油饼中蛋白质含量较高,可作为优质肥料,经脱毒后也可作为饲料~([3]).     

重金属铜、锌、镉复合胁迫对麻疯树幼苗生理生化的影响

该研究以Cu~(2+)、Zn~(2+)、Cd~(2+)单一胁迫为对照,探讨不同浓度的Cu~(2+)、Zn~(2+)、Cd~(2+)复合胁迫对麻疯树幼苗生理生化指标的影响。结果表明:随着Cu~(2+)、Zn~(2+)、Cd~(2+)浓度的增加,麻疯树幼苗叶片中的蛋白质(Pro)、丙二醛(MDA)含量均逐渐增加,其叶片叶绿素含量随着Zn~(2+)胁迫浓度的增加呈现出先降后升的趋势,在中等浓度(100 mg·L-1)的Zn~(2+)胁迫时含量最低、随着Cu~(2+)胁迫浓度的增加叶绿素含量先升高后降低,在Cu~(2+)浓度为200 mg·L-1时含量最高,达到1 200 mg·g-1FW; Cd~(2+)胁迫对叶绿素含量和根系活力无明显影响。根系活力在Zn~(2+)浓度为100 mg·L~(-1)时最强,随着Cu~(2+)浓度的增加而减弱。低浓度的Cu~(2+)、Zn~(2+)、Cd~(2+)对过氧化物酶活性和可溶性糖含量都具有促进作用。Cu~(2+)、Zn~(2+)、Cd~(2+)复合胁迫时对可溶性蛋白、叶绿素和丙二醛含量均无明显影响,随着复合胁迫时浓度的增加,可溶性糖含量和根系活力先增后减。这表明麻疯树对三种重金属的胁迫具有一定的抗性,过高浓度的胁迫会影响麻疯树幼苗生理生化的一些指标,但是麻疯树可以通过自身的防御系统使伤害降到最小。此外,重金属复合胁迫可以在一定程度上减轻单一胁迫对麻疯树幼苗造成的毒害作用。     

麻疯树curcin 启动子的分离及其在转基因烟草中的功能分析

核糖体失活蛋白是一类可以使核糖体28S rRNA的保守a茎环结构域脱嘌呤的蛋白质。麻疯树毒蛋白(curcin)前体基因编码麻疯树胚乳I 型核糖体失活蛋白。从麻疯树基因组中克隆得到其5'侧翼区0.6 kb长的片段, 将该片段插入pBI121载体置换其中的CaMV 35S启动子并在相应的转基因烟草中检测报告基因GUS的表达情况。经过GUS活性检测分析发现, 该0.6 kb长的片段能够启动报告基因在种子中的表达, 并且其在种子不同发育阶段的表达活性存在差异。同时, GUS组织化学染色定位分析表明, 在双子叶植物中该启动子片段是胚乳特异性表达的, 它从心形胚时期开始持续地在烟草的胚乳中发挥启动活性。     

麻疯树核糖体失活蛋白基因的克隆和表达

麻疯树(Jatropha curcas L.)核糖体失活蛋白(curcin)是存在于麻疯树种子中的一种毒性较强的蛋白,它与蓖麻毒蛋白和相思子毒蛋白的性质相似,属Ⅰ型核糖体失活蛋白。从麻疯树种子中分离得到一种分子量为28.2kD的蛋白质,其对无细胞系统中蛋白质合成的抑制活性较强,IC_(50)为(0.19±0.01)nmol/L,具有RNA N-糖苷酶活性。依据curcin的N端部分氨基酸设计简并引物,通过RT-PCR和5′-RACE技术从未成熟种子总RNA中克隆到curcin全长cDNA序列。该cDNA全长由1 173个碱基组成,包含一个编码293个氨基酸的前体蛋白,前42个氨基酸为信号肽。推测的多肽序列与测定的蛋白质N端序列相同,与多种已发表的Ⅰ型核糖体失活蛋白和Ⅱ型核糖体失活蛋白的A链有一定的同源性。将curcin的编码区与表达载体pQE-30相连后,转入大肠杆菌(Escherichia coil)M15菌株中得到了有效的表达。将表达的融合蛋白纯化后发现,它具有抑制无细胞系统蛋白质合成的能力。     

麻疯树毒蛋白curcin2基因的两个原核表达载体的构建和表达

从麻疯树基因组中扩增到毒蛋白curcin2基因成熟肽编码区,成功构建了原核表达载体pET-C和pQE-C,并转化大肠杆菌获得转化子PCB和PCM。在不同温度、不同浓度IPTG和不同诱导时间下,PCB都没有curcin2重组蛋白产生,而PCM能表达出curcin2,主要以包涵体形式存在。诱导温度较低时,延长诱导时间有可溶性的curcin2产生。在文章的实验中,PCM表达curcin2的优化条件为:温度16℃,IPTG1mmol·L-1,时间16￣24h。     

麻疯树核糖体失活蛋白基因的克隆和表达

麻疯树(Jatropha curcas L.)核糖体失活蛋白(curcin)是存在于麻疯树种子中的一种毒性较强的蛋白,它与蓖麻毒蛋白和相思子毒蛋白的性质相似,属Ⅰ型核糖体失活蛋白.从麻疯树种子中分离得到一种分子量为28.2 kD的蛋白质,其对无细胞系统中蛋白质合成的抑制活性较强,IC50为(0.19±0.01)nmol/L,具有RNA N-糖苷酶活性.依据curcin的N端部分氨基酸设计简并引物,通过RT-PCR和5'-RACE技术从未成熟种子总RNA中克隆到curcin全长cDNA序列.该cDNA全长由1 173个碱基组成,包含一个编码293个氨基酸的前体蛋白,前42个氨基酸为信号肽.推测的多肽序列与测定的蛋白质N端序列相同,与多种己发表的Ⅰ型核糖体失活蛋白和Ⅱ型核糖体失活蛋白的A链有一定的同源性.将curcin的编码区与表达载体pQE-30相连后,转入大肠杆菌(Escherichia coil)M15菌株中得到了有效的表达.将表达的融合蛋白纯化后发现,它具有抑制无细胞系统蛋白质合成的能力.     

不同种源麻栎种子形态特征和营养成分含量的差异及聚类分析

以27个种源的麻栎(Quercus acutissima Carr.)种子为实验材料,对不同种源间麻栎种子的形态特征和营养成分含量的筹异性进行分析比较.结果表明,不同种源间麻栎种子的长度、宽度和种子百粒重以及可溶性糖含量、淀粉含量和蛋白质含量差异极显著.麻栎种子的长度和宽度的变化幅度分别为1.69～2.26 cm和1.06～2.50 cm;种子百粒重的变化幅度为83.55～637.38 g;种子可溶性糖含量、淀粉含量和蛋白质含量的变化幅度分别为0.051～0.105 mg·g-1、0.278%～0.471%和21.502～34.696 mg·g-1.麻栎种子百粒重与种子的长度、宽度及可溶性糖含量间显著正相关,种子的长度与可溶性糖含量也显著正相关.主成分分析结果显示,影响麻栎种子品质的主要性状是种子的长度、宽度及种子百粒重.通过聚类分析,将27个麻栎种源划分为3组,其中第1组包括江苏下蜀、安徽太湖、安徽黄山、安徽休宁、安徽滁州、安徽潜山、浙江开化、浙江富阳和山东蒙阴等9个种源,种子品质较好,为优良种源.     

影响农杆菌介导的麻疯树基因转化因素的研究(简报)

麻疯树(Jatropha curcas)为大戟科麻疯树属植物,是一种多年生木本油料植物。原产美洲,广泛分布于热带亚热带地区。因其种子含油量高达 60％,可作生物燃料之用,麻疯树是目前正在被开发利用的重要能源植物之一。除此之外,麻疯树的种子含有多种活性成分,有着重要的农药和医药价值;其生长能耐干旱贫瘠,可用于荒山造林。因此,麻疯树是一种具有多种用途、重要经济价值和学术研究价值的植物。目前对其研究多停留在植物组织培养、植物化学、毒理学、种植业方面等。     

Isolation and Characterization of a Curcin Promoter from <Emphasis Type="Italic">Jatropha curcas</Emphasis> L. and Its Regulation of Gene Expression in Transgenic Tobacco Plants

Ribosome-inactivating proteins (RIPs) represent those proteins that universally depurinate conserved α-sarcin loops of large rRNAs. In this study, a 0.6-kb fragment of a 5′ flanking region preceding a curcin gene, encoding a type I RIP curcin, of Jatropha curcas L. endosperm was cloned, and its regulation of expression of the β-glucuronidase (GUS) reporter gene was investigated in transgenic tobacco. Analysis of GUS activities showed that the 0.6-kb flanking fragment of the curcin gene was sufficient to drive the GUS reporter gene expression in tobacco seed. The activity of this flanking fragment was analyzed at different stages of seed development. Histochemical localization of GUS activity indicated that the promoter was specifically active in the endosperm tissue of the dicotyledonous tobacco embryo. Moreover, this activity was first initiated at the heart-shaped embryonic stage during seed development.     

Somatic embryogenesis in Jatropha curcas Linn., an important biofuel plant

Jatropha curcas L. is one potential source of non-edible biofuel-producing energy crop. Its importance also lies in its medicinal properties. The species is primarily propagated through heterozygous seeds, and thus the seed oil content varies from 4 to 40%. Moreover, due to its perennial nature, seed setting requires 2 to 3 years time. The seed viability and rate of germination are low, and quality seed screening is another laborious task; thus, seed propagation alone cannot provide quality planting material for sustainable use. Somatic embryogenesis, a powerful tool of plant biotechnology for faster and quality plant production has been successfully applied to regenerate plants in Jatropha curcas for the first time. Embryogenic calli were obtained from leaf explants on MS basal medium supplemented with only 9.3 μM Kn. Induction of globular somatic embryos from 58% of the cultures was achieved on MS medium with different concentrations of 2.3–4.6 μM Kn and 0.5–4.9 μM IBA; 2.3 μM Kn and 1.0 μM IBA proved to be the most effective combination for somatic embryo induction in Jatropha curcas. Addition of 13.6 μM adenine sulphate stimulated the process of development of somatic embryos. Mature somatic embryos were converted to plantlets on half strength MS basal medium with 90% survival rate in the field condition. The whole process required 12–16 weeks of culture for completion of all steps of plant regeneration. This protocol of somatic embryogenesis in Jatropha curcas may be an ideal system for future transgenic research.     

Genetic variability,character association and divergence studies in <Emphasis Type="Italic">Jatropha curcas</Emphasis> for improvement in oil yield

Key message

Variability in traits of 15, diverse 6-year-old candidate plus trees of Jatropha curcas was determined to identify the best gain heritable traits correlating with oil yield for Jatropha improvement.

Abstract

Study was carried out on 15 6-year-old candidate plus trees of Jatropha curcas adapted on semi-arid wasteland with an objective to assess variation in morphological, physiological and oil quality characters. Heritable and non-heritable components of the total variability of the characters were determined by genotypic (GCV) and phenotypic (PCV) co-efficient of variation, heritability and genetic advance (GA) and the best gain traits for Jatropha improvement through selection and breeding were assessed. Further, association among the traits were assessed and germplasm were separated into different clusters. Significant variation was found among the different genotypes for all the characters. The photosynthetic and transpiration rate correlated with oil content, seed and oil yield. The chlorophyll pigments correlated positively with the photosynthetic rate and oil content. The seed oil content varied considerably from 27.68 % (JCN01) to 37.49 % (JCN14) and had high heritability, but it had low PCV and GCV and moderate GA. The oil yield plant^?1 had high genetic variability and varied significantly from 0.07 (JCN15) to 0.47 kg plant^?1 (JCN09/IC 565733). Though the different fatty acids differed significantly with different germplasm and also had high heritability, they had low PCV, GCV and GA. Seed weight, fruit weight, seed weight fruit^?1 and seed yield plant^?1 strongly correlated with oil yield and had moderate to high GCV, PCV, coupled with high heritability and GA. Germplasm were separated into four distinct clusters with a maximum inter distance found between cluster II and IV, and minimum between cluster I and III. The study helped to identify the superior germplasm among diverse genotypes of J. curcas that can serve as parents with desirable characters like high oil yield, low stomatal conductance and high water use efficiency for further breeding purposes.

     

Genetic variability, heritability and genetic advance of growth and yield components of Jatropha (Jatropha curcas Linn.) genotypes

A total of 113 Jatropha curcas clonal accessions collected from different regions of India were studied to quantify the magnitude of genetic variability present in the test population and to identify important yield-attributing characters useful for developing high-yielding Jatropha cultivars. High heritability was observed for fruits per plant, seeds per plant, 100-seed weight, seed/kernel (S/K) ratio and kernel oil percentage coupled with high genetic advance suggesting that the accessions can be considered improvement. The significant positive association of seed oil content (%) with 100-seed weight suggested the effectiveness of indirect selection for seed oil content through 100-seed weight. Accessions 76, 120, 29, 86 and 84 showed above average higher values for all yield attributes (viz. fruit and seed yield, 100-seed weight, S/K ratio and oil content) suggesting these as best out of the test accessions. Accessions showing higher values for one or the other yield attributes could be selected as parents for further improvement.     

Tomato sugar transporter genes associated with mycorrhiza and phosphate

A new kind of ribosome-inactivating protein (curcin 2), induced by several different kinds of stress from Jatropha curcas leaves, under the control of the CaMV (cauliflower mosaic virus) 35S promoter, was introduced into the tobacco genome by Agrobacterium tumefaciens-mediated transformation method. The curcin 2 protein was only detected in the transgenic tobacco plantlets transformed with the cur2p fragment (coding premature curcin 2 protein), but not in the plantlets with the cur2m fragment (coding mature curcin 2 protein). The T1 population of the transgenic lines shows an increased tolerance to tobacco mosaic virus (TMV) and a fungal pathogen Rhizoctonia solani by delaying the development of systemic symptoms of TMV and reducing the damage caused by the fungal disease. The increases of the tolerances correspond to the curcin 2 level in the transgenic plants.     

Variability in reproductive traits in Jatropha curcas L. accessions during early developmental stages under warm subtropical conditions

Variability in floral, fruit, and seed characteristics, and oil content of 15 accession of Jatropha curcas during early development were assessed during two flowering periods in south Florida subtropical climate. The two flowering periods had leaf flushing in March. Field evaluation using 18 quantitative traits showed significant variation among accessions. The number of female flowers and female : male flower ratio ranged from 1 to 15 and 1 : 8.8 to 1 : 67.8, respectively. Fruit set by natural pollination was 89 and 66% during the first (1st) and second (2nd) flowering periods, respectively. A higher number of female‐type inflorescences were observed during summer. There were significant differences in seed traits, except for number of seeds per fruit. Accession TREC 31 had the highest individual seed dry weight and 100‐seed weight (0.83 g and 79.7 g, respectively). The oil content varied from 19.30% to 35.62%. Seed dry weight had positive correlation with seed fresh weight, seed length, seed thickness, seed width, and 100‐seed weight, but negative correlation with oil content. Based on the cluster analysis using 15 morphological traits, jatropha accessions were grouped into five main clusters and accessions from different geographic regions grouped together in a cluster. Principal component analyses (PCA) revealed morphological variation. The first three components explained 73.5% of the total variation and seed dry weight, 100‐seed weight, total flowers per inflorescence, male flowers per inflorescence and fruit set can be used to distinguish accessions. The PCA also indicated that flowering traits were more influenced by seed origin while seed traits were affected by flowering spans. Although evaluations were performed in plants during the juvenile phase, accessions TREC 31 and TREC 55 had superior averages for almost all characters evaluated. These results provide a preliminary assessment of the high variability in jatropha accessions evaluated and their potential for use in breeding and genetic improvement programs.     

金沙江干热河谷滇榄仁种子扩散与种子库特征研究

采用随机取样和跟踪调查的方法对滇榄仁(Terminalia franchetii)种子扩散规律、空间分布特征以及种子库动态进行了系统研究。结果表明,滇榄仁种子扩散受到较强的风力作用影响,具有明显的方向性,扩散距离是其树高的1.6倍以上。不同群落类型中,滇榄仁的土壤种子库存在较大差异,但不同时间段的变化趋势基本一致,即在雨季(6月、8月和10月)数量相对较少,12月到翌年4月,数量逐渐增加。滇榄仁植冠种子库从当年10月开始缓慢脱落,到翌年3月达到脱落高峰(15%以上种子此时脱落),然后再次进入缓慢脱落期,翌年6、7月的种子脱落速率最低,但直到翌年10月仍有部分种子(约15%左右)存在于植株冠层。总体来说,不同类型群落中,滇榄仁幼苗更新不良的原因不应是缺乏种源,可能与种子质量以及群落环境等影响群落天然更新的其它因子存在较大关联。同时,滇榄仁土壤种子库数量与植冠种子库的脱落过程存在紧密联系,而植冠种子脱落可能与该区域气候因子存在着较大依存关系,其中,与风力可能存在着正相关,与降雨量和空气相对湿度存在着负相关,这可能是滇榄仁长期以来对干热河谷环境的一种适应特征。     

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。