麻疯树(Jatropha curcas L．)幼苗生长和光合作用对盐胁迫的响应

为探讨盐胁迫对麻疯树幼苗生长和叶片光合生理的影响,在温室不同浓度NaCl处理下,对生长、光合色素含量、净光合速率Pn等光合参数、叶片MDA含量和电解质相对外渗率EL进行测定。结果表明:①25～50mmol·L-1NaCl处理促进幼苗生长,尤其是50mmol·L-1NaCl处理,鲜重较对照显著增加44.3%,100～150mmol·L-1NaCl对生长影响不大,200～250mmol·L-1NaCl使生长受抑制,鲜重较对照分别显著下降39.3%和70.2%。②50mmol·L-1NaCl时,Pn较对照显著增加17.2%,100～150mmol·L-1NaCl时与对照无显著差异,200～250mmol·L-1NaCl时分别比对照显著下降73.2%和77.9%,Gs和Tr呈相同趋势,MDA含量、EL则相反。③叶绿素Chl在25～50mmo.lL-1NaCl时递增,100～250mmol·L-1NaCl时递减,但Chla/Chlb值变化小;类胡萝卜素Car在25mmol·L-1NaCl时显著增加,100mmol·L-1NaCl后缓慢下降,但Car/Chl值呈上升趋势。④25～150mmo.lL-1NaCl时幼苗水分利用效率WUE与对照无显著差异,200～250mmol·L-1NaCl时比对照显著下降。综上所述,25～50mmol·L-1NaCl时,麻疯树幼苗通过增加气孔开张、增强光合膜等细胞膜稳定性和膜功能,使Pn显著增加,促进植株生长和提高耐盐性,100～150mmo.lL-1NaCl时,通过稳定的WUE使Pn下降不显著,生长受影响小,具有较好的耐盐性,200～250mmo.lL-1NaCl时,光合膜等生物膜功能减弱,使Pn显著下降,幼苗诱导活性氧清除系统并通过减小生长来提高耐盐性,具有较好的耐盐适应性。150mmol·L-1NaCl以下是麻疯树幼苗生长的适宜浓度。

金沙江干热区麻疯树(Jatropha curcas L.)杂交育种技术的初步研究

对金沙江干热区麻疯树生物特性、开花特性、结实特性进行观测,并开展不同地理种源麻疯树杂交育种试验。结果表明,麻疯树在本区域生长发育较好,可持续开花结果至1月初,雌雄花比例可达1∶10左右。通过麻疯树杂交育种试验,研究了麻疯树雌雄花分辨、雌花可授粉时间、套袋材料及去袋时间等特性。通过对金沙江干热区麻疯树主要特性和杂交育种技术试验研究,有利于提高育种效果,加速育种进程,提高育种工作的预见性。

麻疯树(Jatropha curcas L.)高含油量单株无性系筛选评价

利用初步筛选出的种仁含油量大于65%的10个麻疯树地理种源单株,以种仁含油量为目标,在元谋干热河谷区,经过3次无性繁殖复选,并对其种仁含油量进行测定评价。结果表明:由于生长环境的改变,10个单株材料的种仁含油量都有不同程度的下降,但来自元谋物茂乡、双柏大庄乡和云南永仁县的3个单株材料种仁含油量下降幅度较小,且连续表现稳定,可作为进一步品种选育和生产使用的无性系材料。

Changes of Water Status and Different Responses of Osmoregulants in Jatropha Curcas L. Seedlings to Air-drought Stress

[Objective] The study aimed to investigate the changes of water status and different responses of osmoregulants during air-drought stress,to better understand mechanisms of drought resistance in Jatropha Curcas L. [Methods] The 12-day-old J. curcas seedlings were held in a climate chamber at 25/20 ℃（day/night）,16 hours illumination,and 75% of relative humidity for air-drought treatment,and the changes of water potential,osmotic potential and the content of soluble sugar,proline,betaine were measured. [Results] Water potential and osmotic potential in leaves of J. curcas seedlings dropped significantly,pressure potential lost during air-drought stress,and the contents of osmoregulants soluble sugar,proline and betaine rose significantly to different extent in the leaves and stems. [Conclusion] Osmoregulants in the leaves and stems respond differently to air-drought stress,and in general leaves are much more responsive to the drought than stems of J. curcas seedlings.

Distribution and development strategy for Jatropha curcas L. in Yunnan Province, Southwest China

Yunnan Province is the main distributing area ofJatropha curcas L. This plant is abundant in several drainage areas of the dry-hot, dry-warm and sub-humid valleys in the south subtropical area of Yunnan Province. The seeds that were picked from trees blossoming between April and May and fructifying between September and October will have large seed yield and fine quality. For developing bio-diesel stock forest ofJ. curcas in areas with adaptive climate, seeding measures for afforestation should be taken and techniques on breeding, fast-growing, and high-yielding plantation cultivation are very important.

<i>In</i><i>Silico</i>Mining of EST-SSRs in Jatropha curcas L. towards Assessing Genetic Polymorphism and Marker Development for Selection of High Oil Yielding Clones

In recent years, Jatropha curcas L. has gained popularity as a potential biodiesel plant. The varying oil content, reported between accessions belonging to different agroclimatic zones, has necessitated the assessment of the existing genetic variability to generate reliable molecular markers for selection of high oil yielding variety. EST derived SSR markers are more useful than genomic markers as they represent the transcriptome, thus, directly linked to functional genes. The present report describes the in silico mining of the microsatellites (SSRs) using J. curcas ESTs from various tissues viz. embryo, root, leaf and seed available in the public domain of NCBI. A total of 13,513 ESTs were downloaded. From these ESTs, 7552 unigenes were obtained and 395 SSRs were generated from 377 SSR-ESTs. These EST-SSRs can be used as potential microsatellite markers for diversity analysis, MAS etc. Since the Jatropha genes carrying SSRs have been identified in this study, thus, EST-SSRs directly linked to genes will be useful for developing trait linked markers.

Growth and Transpiration of <i>Jatropha curcas</i>L. Seedlings under Natural Atmospheric Vapour Pressure Deficit and Progressive Soil Drying in Semi-Arid Climate

During the last decade, Jatropha curcas L. (J. curcas) has gained much attention worldwide as biofuel crop. Although its cultivation is promoted in the Sahel, there is a surprising lack of data on its water use regulation and growth in this region. Here, we investigated, in semi-controlled conditions, leaf transpiration and growth of six accessions of J. curcas at seedling stage under natural changing in vapour pressure deficit (VPD) and progressive soil drying in Senegal. The experimental layout was a randomized complete bloc design and after 3 months of growth arranged to a split-plot at the implementation of water stress to facilitate irrigation. Under well water condition, there was no significant difference between accessions for leave transpiration that was positively correlated to VPD with high values recorded between 13 h and 14 h pm. Accessions of J. curcas used in this study showed closed thresholds at which transpiration declined except accession from Ndawene that threshold was lower (0.30). There is no significant difference between accessions for growth during the experimentation period. In 3 months, we recorded 23.57 g for the aboveground dry biomass and seedlings had about 14 leaves and 24.3 cm height. Positive linear correlation was recorded between aboveground biomass and root dry weight (p J. curcas might need complement irrigation for a better growth of seedlings especially during the dry season.

Synthesis of biodiesel from Jatropha curcas L. seed oil using artificial zeolites loaded with CH3COOK as a heterogeneous catalyst

An environmentally benign process was devel-oped for the transesterification of Jatropha curcas L. seed oil with methanol using artificial zeolites loaded with potassium acetate as a heterogeneous catalyst. After calcination for 5 h at 823 K, the catalyst loaded with 47 wt.% CH3COOK exhibited the highest efficiency and best catalytic activity. The easily prepared cata-lysts were characterized by means of X-ray dif-fraction and IR spectroscopy, as well as Hammett indicator titration. The results revealed a strong dependence of catalytic activity on ba-sicity. The optimum reaction conditions for transesterification of J. curcas oil were also in-vestigated. The methyl ester content in the bio-diesel product exceeded 91% after 4h reaction at reflux temperature in the presence of 2% solid catalyst and no water washing process is needed during workup.

Biodiesel Production from Crude Jatropha curcas L.Oil with Trace Acid Catalyst

Biodiesel produced from crude Jatropha curcas L.oil with trace sulfuric acid catalyst(0.02%-0.08% oil) was investigated at 135-184 ℃.Both esterification and transesterification can be well carried out simultane-ously.Factors affecting the process were investigated,which included the reaction temperature,reaction time,the molar ratio of alcohol to oil,catalyst amount,water content,free fatty acid(FFA) and fatty acid methyl ester(FAME) content.Under the conditions at 165 ℃,0.06%(by mass) H2SO4 of the oil mass,1.6 MPa and 20:1 methanol/oil ratio,the yield of glycerol reached 84.8% in 2 hours.FFA and FAME showed positive effect on the transesterification in certain extent.The water mass content below 1.0% did not show a noticeable effect on trans-esterification.Reaction kinetics in the range of 155 ℃ to 175 ℃ was also measured.

One-step process for production of biodiesel from Jatropha curcas L. seeds： Optimization using Response Surface Methodology （RSM）

Jatropha curcas L. （JCL） seeds were extracted and transesterified in-situ using supercritical methanol extraction in the absence of catalyst at different temperatures （200-280℃） and pressures （8-12 MPa）, and at a fixed reaction time of 30 min with seeds-to-methanol ratio of 1：40 w/v. Design of experiment approach using five-level-two-factors design of Response Surface Methodology （RSM） was used to observe the effect of two independent variables i.e. temperature and pressure and the percent of biodiesel yield which required 13 runs. For optimization of the variables, Central Composite Rotatable Design （CCRD） was used for regression analysis and analysis of variance （ANOVA）. The optimize conditions suggested by RSM were at T = 280℃ and P = 12.04 MPa. The predicted and experimental biodicsel yields were found to be 56.8% and 59.9%, respectively, with relatively small deviation errors of 1.59%.

Gene cloning,expression analysis of JcACP (Acyl Carrier Protein) in Jatropha curcas L. and its prokaryotical expression

Objective To clone the ACP(acyl carrier protein)gene in Jatropha curcas L.,a potential anti-tumour and anti-fungal plant.And to determinate the expression of ACP in Jatropha curcas L.Methods A cDNA clone encoding ACP(acyl carrier protein)was isolated from Jatropha curcas L.endosperm cDNA library by random sequencing.The expression of ACP gene was investigated by semi-quantitative RT-PCR in leaves,stems and seeds of J.curca.The expression of ACP was also investigated in germinating seeds.The fragment encoding ACP protein in J.curca.was inserted into a prokaryotic expression vector pET28a(+).The gene was overexpressed in E.coli BL21 to produce abundant protein.Immunohistochemical analysis was used to detect the expression of ACP in different tissues of J.curca.Results The cDNA sequence was 806 bp in length and the ORF was 393 bp.The predicted molecular weight of the putative protein was 14.4 kD,pI=5.2.It contained a 4'-phosphopantetheine-binding motif.This prosthetic group can be combined with Serine of ACP protein.Semi-quantitative RT-PCR analysis showed that ACP gene was expressed in leaves,stems and seeds of J.curcas.The expression level of ACP was the highest in seeds and it was not detected in roots.After seeds germinated,the expression level of ACP in seeds increased progressively and reached a peak at 96 h.After induced by IPTG,SDS-PAGE analysis showed that the ACP protein of 20 kD was expressed.Immunohistochemical analysis showed that ACP specifical expressed abundantly in embyo of the seeds,and it was not detected in roots and the emdosperm while expressed in leaves and stems.Conclusions A cDNA clone encoding ACP which had all the typical characteristics of ACPs was isolated.It was expressed successfully in E.coli.The results of semi-quantitative RT-PCR analysis and immunohistochemical analysis were very similar,which showed that the expression of ACP in J.curcas.was abundant in seeds.The results indicated the expression related to the high metabolism.

Effects of Aluminum Toxicity on Physiological Indices of Jatropha curcas L.Seedling

Jatropha curcas was taken as the test material,6 concentrations including 0,25,50,100,200 and 400μmol/L AlCl3,plus 3 time gradients including 7,14 and 21 d,were set to study the effects of Al^(3+)stress on the antioxidant system of Jatropha curcas L.seedling.The results showed that with the Al^(3+)treatment being applied,protein content increased first,then decreased and finally increased with the increase of Al^(3+)concentration;the soluble sugar content increased first and then decreased with the increase of Al^(3+)concentration.Under low concentration of Al^(3+)treatment,Pro content,MDA content and POD activity of Jatropha curcas L.seedling leaves changed a little,while under high concentration of Al^(3+)treatment,Pro and MDA content of Jatropha curcas L.seedling leaves rapidly accumulated,POD activity increased and they showed a trend of increase with the increase of Al^(3+)concentration;From the perspective of Al^(3+)stress time,protein content,soluble sugar content,MDA content and POD activity increased with stress time being prolonged,while Pro content decreased with stress time being prolonged.These results indicated that the leaves of Jatropha curcas L.seedlings had certain self-protection and remediation abilities under Al^(3+)stress.

Cloning and expression analysis of HXK1 gene from Jatropha curcas L.

HXK(Hexokinase)gene family and the role of JcHXK1 in Jatropha curcas L.were explored.Totally 4 HXK genes JcHXK1,JcHXK2,JcHXK3 and JcHKL1 were identified and complete ORF of JcHXK1 was cloned.Functional domain,phylogenetic evolution and low-temperature expression characteristics were analyzed.Results showed that full-length JcHXK1 cDNA was 1497 bp,encoding 498 amino acids with molecular weight of 53.81 kDa and pI of 5.03.Further phylogenetic evolutionary analysis demonstrated HXK1 protein was clustered into 6 small branches and 2 large branches.Sequence alignment showed that HXK1 contained several conserved glycine residues and hydrophobic channels.Prokaryotic expression vector of JcHXK1 was constructed and 12%SDS-PAGE detection showed that it was highly expressed in E.coli.These research was expected to lay a foundation for further gene functional verification and cold signal transduction mechanism for HXK1 in Jatropha curcas L.

麻疯树外植体愈伤组织中毒蛋白的Western-blot鉴定

The proteins from endosperm, root, stem, leaf, leafstalk of Jatropha curcas L. and their calli were hybridized to the seed toxin protein (curcin) of Jatropha curcas L. by western blot. The result showed that the curcin was specifically expressed in the endosperm and its calli, while it was not detected in root, stem, leaf, and leafstalk of Jatropha curcas L. and their calli . This study indicated that calli induced from endosperm can be used to produce curcin.

麻疯树提取物活性物质及抗菌作用研究

使用乙醇—乙酸(1∶1)回流提取方法,获得麻疯树枝叶中的香豆素类化合物,并使用液相色谱—质谱法(LC-MS)对提取物进行定性及定量分析,通过滤纸片法探究了麻疯树中的香豆素类化合物对两种细菌的体外抑菌效果,并通过肉汤微量稀释法和生长曲线法测定麻疯树提取物(Jatropha curcas Extraction,JE)对金黄色葡萄球菌和大肠杆菌的最小抑菌浓度(MIC)和最低杀菌浓度(MBC)。结果表明:提取的香豆素类化合物中,主要成分为佛手柑内酯(Bergapten),相对含量为18.66%,在麻疯树化学成分研究中首次发现;同时还发现黄芩素(Baicalein)和野漆树苷(Rhoifolin),相对含量为2.76%和0.16%;麻疯树香豆素类化合物对金黄色葡萄球菌和大肠杆菌均有明显抑制效果;JE对金黄色葡萄球菌和大肠杆菌的MIC分别为1.56 mg/mL和3.125 mg/mL,对二者的MBC均为6.25mg/mL。

多效唑浸种对NaCl胁迫麻疯树幼苗的生长调节效应

以南油1号麻风树幼苗为材料,通过盆栽试验研究了200 mmol·L^-1NaCl胁迫处理对不同浓度（0～1 000 mg·L^-1）多效唑（PP333）浸种麻疯树幼苗生长和光合作用的影响.结果显示：在盐胁迫下,不同浓度的PP333浸种处理均显著降低麻疯树植株株高,但是均显著提高了盐胁迫下株高相对生长速率,同时提高了其干物质积累速率、根含水量、根冠比、叶绿素含量和净光合速率;并提高了幼苗根、叶的K^＋含量,降低根、叶的Na^＋和Cl^-含量,从而提高根、叶的K^＋/Na^＋比率.研究表明,PP333浸种能缓解盐胁迫下麻疯树幼苗的失水程度和光合色素的下降幅度,有效改善其光合作用效率,同时维持体内的离子平衡,从而减轻盐胁迫伤害,促进植株生长,并以600 mg·L^-1PP333浸种效果最好.

HPLC法测定麻疯树不同部位两种二萜的含量

本文建立了两种麻疯树二萜麻疯树酚酮B和Curcusone B的含量测定方法。采用Alltima C18色谱柱(4.6mm×250mm,5μm),流动相为甲醇-水(70∶30),流速为1.0 mL·min-1,紫外检测波长为254 nm。实验结果表明:麻疯树酚酮B和Curcusone B的线性范围分别为2.26-113.0μg·mL-1(r=0.9998)和2.06-103.0μg·mL-1(r=0.9997),平均回收率分别为96.7%(RSD=1.71%)和99.3%(RSD=1.69%)。所建立的方法简便、快速、准确,重现性好,成功用于麻疯树不同部位两种二萜的含量测定。结果显示,麻疯树根皮中二萜含量最高,麻疯树酚酮B和Curcusone B达到了0.31%和0.13%。茎皮次之,分别为0.012%和0.020%。木质部分和树叶中未检测到两种二萜的存在。

麻风树JcHDZ28基因克隆与功能分析

HD-Zip家族基因与植物的生长和对环境胁迫的抗性密切相关,被认为是作物改良的关键因子。在本研究中,我们通过RT-PCR技术从麻风树中克隆了1个HD-Zip家族基因,命名为JcHDZ28。JcHDZ28基因包含1个882 bp的开放阅读框,编码1个含有293个氨基酸的蛋白质。氨基酸序列分析结果表明,JcHDZ28含有高度保守的同源结构域和亮氨酸拉链(LZ)基序。表达模式分析结果表明,JcHDZ28基因在种子中的相对表达量最高,且盐胁迫下调该基因的表达。亚细胞定位分析结果表明,JcHDZ28基因编码1个核定位蛋白。过表达JcHDZ28基因增加了转基因拟南芥对盐胁迫的敏感性,且在盐胁迫条件下,转JcHDZ28基因拟南芥叶片脯氨酸含量显著低于野生型拟南芥,相对电导率显著高于野生型拟南芥,非生物胁迫相关基因在转JcHDZ28基因拟南芥中的表达也显著低于在野生型拟南芥中的表达。本研究结果可以为进一步阐明HD-Zip转录因子基因JcHDZ28在麻风树生长发育和响应非生物胁迫中的功能提供理论依据。

小桐子果壳杀虫活性部位的分离及其HPLC-MS分析

为寻找小桐子果壳的杀虫活性物质,以小桐子果壳乙醇粗提物的正丁醇萃取相为实验材料,对其采用硅胶柱反复柱层析,同时对各阶段分离产物进行活性追踪,筛选出具有杀虫作用的活性部位WD1,最后采用HPLC-MS分析,初步鉴定出活性部位主体成分。结果表明,WD1活性组分在浓度为8g·L-1时,对桃蚜(Myzus persicae)和菜青虫(Pieris rapae)的24h(72h)校正死亡率均大于70%,显示出极高的毒杀活性。通过对WD1进行HPLC-MS分析,鉴定了含量较高的7个化合物,为总量的80.72%,初步鉴定为2a,3a,24-三羟基-12-20(30)-二烯-28-乌索酸、环辛肽B、牡荆素、松香烷二萜、麻疯树酚酮A、Curcusone A或Curcusone B。

超表达JcFT基因对烟草农艺性状的影响

FT(Flowering locus T)基因编码的成花素蛋白,也称蛋白激素,在植物中超表达该基因往往可以显著提早开花,是创制早熟植物种质资源的重要潜在基因之一,影响植物生长发育。本研究以烟草为材料,分析了FT对烟草多个农艺性状的影响。结果表明,JcFT(FT of Jatruopha curcas L.)超表达在提早烟草开花的同时,显著改变了烟草根、茎、叶的大小和形态,而花的结构没有显著改变。