Cloning and Expression of Curcin, a Ribosome-Inactivating Protein from the Seeds of Jatropha curcas

Curcin, a ribosome-inactivating protein with a molecular weight of about 28.2 kD, which strongly inhibits the protein synthesis in rabbit reticulocyte lysate system with an IC50 value of about (0.19 +/- 0.01) nmol/L, was purified from the seeds of Jatropha curcas L. The protein has the activity of rRNA N-glycosidase. Degenerate primers were designed based on the N-terminal partial sequence from purified curcin. The full-length curcin cDNA by RT-PCR and 5'-RACE was cloned. The deduced amino acids sequence indicates that a preprotein with 20 amino acid residues is first translated and then processed to a mature protein with 251 amino acids. The deduced amino acids sequence shares homology of 33% and 57% to those of type I ribosome-inactivating proteins (RIPs) and A chain of type II RIPs, respectively. The sequence encoding mature curcin was integrated into the pQE-30 vector for expression in Escherichia coli strain M15 (pREP4). The purified recombinant curcin was able to inhibit protein synthesis in rabbit reticulocyte lysate system.

对药用及能源植物Jatropha curcas L.的中文名称的建议

目前重要药用及能源植物Jatropha curcas L.的中文名称很多,给生产应用和科研等带来诸多不便。统一命名,特别是在当今信息化时代,对于学术交流和科技推广具有极其重要的意义。根据我国各地的中文名称使用习惯和大众心理,我们建议将Jatropha curcas L.的中文名称统一为"小油桐"。

Study on Establishment of cpSSR Marker Technology and Optimization of Its Reaction System in Jatropha curcas Linn

[ Objective] The aim of this study was to establish the optimum cpSSR-PCR system for Jatropha curcas Linn. [ Method] cpSSR-PCR amplification system for Jatropha curcas Linn influenced by five factors including Taq DNA polymerase, Mg^2＋ , DNA template, dNTP and primer were optimized from several levels. [ Result] The optimum concentration of 20 μl reaction system was 10 × Buffer, 2.00 mmol/L Mg^2＋ , 2 U/μl Taq DNA polymerase, 0.2 mmol/L dNTP, 0.2 μmol/L primer and 35 ng/μl DNA template. [ Conclusion] The optimum annealing temperature for cpSSR-PCR reaction system is 52 ℃, and the cpSSR reaction system is steady and reproducible.

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.

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。

云南麻疯树(Jatropha curcas)资源生态地理分布及评价

对云南省麻疯树的分布区、资源量和粗脂肪含量开展系统调查研究。结果表明:云南麻疯树资源分布在金沙江、澜沧江、怒江和红河流域的260乡(镇),呈零星分布,不同产地种子粗脂肪含量差异较大,单株间亦存在差异;81个种群所采种仁中,永仁县永兴乡种仁粗脂肪含量最高达67.00%,屏边白河最低为34.88%。评述了云南省的小桐子资源状况,并术其资源及利用的价值优势,论述了云南麻疯树的开发利用前景。

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。

甲基磺酸乙酯(EMS)处理对麻疯树(Jatropha curcas L.)萌发种子的生理效应

分析了不同甲基磺酸乙酯(EMS)浓度(0、0.1%、0.5%、1.0%、1.5%和2.0%)和在浓度为1.2%时不同处理时间(0、8、12、16h和20h)对麻疯树萌发种子的生理效应。结果表明,EMS对麻疯树种子的半致死浓度为1.2%,当浓度达到2.0%时,种子受到严重伤害,成苗率为1.67%。用1.2%浓度诱变麻疯树种子,随着处理时间的延长,种子受到的伤害也随之加重,并且产生的矮化植株率也随之大幅度升高,在处理时间大于12h时,得到的植株都是矮化植株,但当处理时间达到20h时种子成苗率为0。由此看来,用EMS诱变麻疯树种子的合适浓度为1.2%,处理时间应选在8～12h范围内。

金沙江干热区麻疯树(Jatropha curcas L.)白粉病危害特征观察

在麻疯树的规模种植区选定代表性的10株无病植株,观察白粉病的危害侵染特征。结果显示:群居的麻疯树极易感染白粉病,在麻疯树的整个生长期都会受到不同程度的危害,8～10月高温高湿条件下是发病的高峰期,11月份以后减缓危害速度,但不停止。结果对麻疯树白粉病预警预报和有效防控措施的制定提供参考依据。

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。

Jatropha curcas L: Phytochemical, antimicrobial and larvicidal properties

Objective: To evaluate antimicrobial activities as well as the phytochemical and lavicidal properties of different parts of Jatropha curcas L.(J. curcas) growing in Mauritius.Methods: Determination of the presence of phytochemicals in the crude plants extracts by test tube reactions. Disc diffusion method and microdilution method were used to detect the antimicrobial sensitivity and activity(minimal inhibitory concentration). The crude solvent extracts were also tested on the larvae of two insects, Bactrocera zonata and Bactrocera cucurbitae(Diptera, Tephritidae).Results: The antimicrobial activities were significantly dependent for the different crude plant extracts on the thirteen microorganisms tested. For the Gram-positive bacteria, the crude ethyl acetate extract was more efficient compared to the Gram-negative bacteria with both solvents being effective. The crude ethyl acetate extract of J. curcas bark and mature seed oil showed the highest efficacy. The highest mortality percentage was observed after 24 h for both Diptera flies with(66.67 ± 2.89)% of Bactrocera cucurbitae larvae killed by ethyl acetate extract of J. curcas bark.Conclusions: This paper compared the different J. curcas plant sections with respect to the effectiveness of the plant as a potential candidate for new pharmaceuticals. The larvicidal effect was also studied in order to demonstrate the dual purpose of the plant.

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.

Detailed investigation of optimized alkali catalyzed transesterification of Jatropha oil for biodiesel production

The non-edible oils are believed to be one of the major feedstock for the production of biodiesel in future.In the present study,we investigated the production of Jatropha oil methyl esters（JOMEs） via alkali-catalyzed transesterification route.The biophysical characteristics of Jatropha oil were found within the optimal range in accordance with ASTM standards as a substitute diesel fuel.The chemical composition and production yield of as-synthesized biodiesel were confirmed by various analytical techniques such as FT-IR,1H NMR,13 C NMR and gas chromatography coupled with mass spectrometry.A high percentage conversion,～96.09%,of fatty acids into esters was achieved under optimized transesterification conditions with 6 ：1 oil to methanol ratio and 0.9 wt% Na OH for 50 min at ～60°C.Moreover,twelve fatty acids methyl esters（FAME） were quantified in the GC/MS analysis and it was interesting to note that the mass fragmentation pattern of saturated,monounsaturated and diunsaturated FAME was comparable with the literature reported values.

麻疯树(Jatropha curcas L.)种子总DNA提取方法的建立和优化

用种子提取DNA可省却培育幼苗过程而加快实验进度,为此建立了麻疯树(Jatropha curcas L.)种子DNA提取方法。针对麻疯树种子富含蛋白质、多酚及多糖等次生物质的特点,基于CTAB法进行优化,在研磨种子时加入抗氧化剂PVP去除多酚,接着用核分离缓冲液去除多糖,再通过酚-氯仿-异戊醇(体积比=25∶24∶1)抽提去除蛋白质。所提取的麻疯树种子总DNA浓度和质量均较高,经琼脂糖凝胶电泳和ISSR-PCR扩增得到了非常清晰的DNA条带。

Floral display and breeding system of Jatropha curcas L.

Plant flowering and breeding characteristics are important for us to understand the reproduction of plant populations. In this paper, we studied the reproduction characteristics of Jatropha curcas in Yuanjiang County （23°36＇1＇4, 101°00＇E）, Yunnan Province. The plant produces flowers in dichasial inflorescences. Normally, the flowers are unisexual, and male and female flowers are produced in the same inflorescence. Only a few male flowers are produced in an inflorescence, and fruits are produced only through pollination between different flowers from the same or different plants. By the treatments of emasculation, bagging and artificial pollination in this experiment, there were few but same fruit set ratios when the inflorescences were emasculated, bagged, or bagged with net, except artificial pollination treatments, which showed that Jatropha curcas could produce fruit through apomixis but not wind pollination. When the inflorescences were unbagged, unemasculated and with free pollination treatments, or bagged, emasculated and with artificial cross-pollination treatments, or unbagged, emasculated and with free pollination treatments, there were many fruits produced. It showed that Jatropha curcas shows outcrossing, is self-compatible, and demanding for pollinators. Normally, the male flowers open first and a few flowers bloom in one day in a raceme. These flowers last a long time in bloom. However, a large number of female flowers open from the third to the fifth day, with some female flowers opening first in a few raceme. This shows a tendency to promote xenogamy and minimize geitonogamy.

Cloning and characterization of a stearoyl-ACP desaturase gene from Jatropha curcas

Using degenerate primers and RT-PCR, RACE techniques, a 1491 bp cDNA segment of stearoyl-acyl carrier protein desaturase （SAD） is cloned from developing seeds of Jatropha curcas L. The segment contains a 1191 bp of complete open reading frame （ORF）. Analysis in the BLAST on NCBI shows that Jatropha curcas SAD （JSAD） gene encodes a protein precursor composed of a signal peptide of 33 amino acids and a mature peptide of 363 amino acids. The homological analysis shows that JSAD has high level of homology both in nucleotide sequence and in amino acid sequence to other plants SADs. The nucleotide and peptide identity of JSAD to Ricinus communis SAD （RSAD） is up to 89% and 96.2% respectively. Molecular modeling of JSAD indicates that its three-dimensional structure strongly resembled the crystal structure of RSAD.

Comparative characterization of Jatropha,soybean and commercial biodiesel

Oil was extracted from seeds of Jatropha Curcas,in high yields(up to 40% by weight).The extracted Jatropha oil was converted in a laboratory reactor to biodiesel by transesterification.Analysis of Jatropha oil and Jatropha biodiesel by GC/MS and GC/SIMDIS showed that Jatropha oil could be readily converted to a biodiesel product through NaOH catalyzed transesterification.The resulting biodiesel has desirable properties such as high cetane number and low flash point,which are major improvements over the properties of commercial biodiesel fuels.

The biomedical significance of the phytochemical, proximate and mineral compositions of the leaf, stem bark and root of Jatropha curcas

Objective: To analyse the phytochemical contents of leaf, stem bark and root of Jatropha curcas(J. curcas) in four solvent extracts and their proximate and mineral compositions. Methods: Standard analytical procedures were used for the determination of phytochemicals, proximate and mineral compositions of the leaf, stem bark and root extracts of J. curcas. Results: Results of the analysis showed the presence of polyphenols, flavonoids, alkaloids, cardiac glycosides, coumarins, saponins, terpenoids, steroids, triterpenoid saponins, carotenoids, phlobatannins and tannins in the leaf, stem bark and root of all the solvent extracts. Flavonoids were present in the highest amount in the ethyl acetate extracts of the leaf(7.35% ± 0.02%), stem bark(4.12% ± 0.01%) and root(3.35% ± 0.02%) followed by polyphenols in the methanol extracts of leaf(4.62% ± 0.02%), stem bark(2.77% ± 0.05%) and root(2.49% ± 0.02%). Poly-acetylated compounds were absent in all the solvent extracts of the leaf, stem bark and root. However, some anti-nutritional agents such as oxalates, phytates and cyanates were present in all the solvent extracts of the leaf, stem bark and root except the ethyl acetate. Phytates were high in the aqueous solvent of the leaf(6.12% ± 0.00%) but low in the stem bark(1.00% ± 0.05%) and root(0.89% ± 0.03%). Proximate composition showed appreciable amounts of total carbohydrate(36.33% ± 0.72%), crude protein(26.00% ± 0.47%) and reducing sugars(5.87% ± 0.14%) in the leaf, while crude fat was more in the stem bark(16.70% ± 0.30%). There was corresponding substantial energy in the leaf [(1 514.77 ± 20.87) kJ /100 g] and stem bark [(907.00 ± 8.52) kJ /100 g]. Moisture and ash contents of the leaf, stem bark and root were within acceptable limits for the use in drugs formulation. The mineral composition showed substantial amounts of important elements such as Fe, Ca, Na, Mg and Zn. Others were P, K and Se. Conclusions: The outcome of this study suggests that the leaf, stem bark and root of J. curcas have very good medicinal potentials, meet the standard requirements for drug formulation and serve as good sources of energy and nutrients except for the presence of some anti-nutritional elements predominant in the leaf.

Isolation of antileishmanial,antimalarial and antimicrobial metabolites from Jatropha multifida

Objective:To investigate the antileishinanial,antimicrobial and antimalarial activities of the pure metabolites from Jatropha multifida used in African ethnomedicine.Methods:The methanolic stem bark extract of Jatropha multifida used in Nigerian folk medicine as remedy against bacterial infections was subjected to column chromatography and HPLC analyses lo obtain three known metabolites,microcyclic lathyrane dilerpenoids(1-3).Structures were confirmed by comparison of 1D and 2D spectral data with literature.Results:The three compounds exhibited inhibition of antileishmanial,antimalarial and antimicrobial actions against the tested organisms with compouds 2 and 3 active against Cryptococcus neoformans at IC_(50)of 82 and 8.7 μg/ml,respectively.Conclusions:The research lends support to the ethnomedicinal use of the plant in combating microbial infections,leishmaniasis and malarial infections.

<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.