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.

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。

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。

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。

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。

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。

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.

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.

Development and Characterization of Eco-Friendly Non-Isocyanate Urethane Monomer from Jatropha curcas Oil for Wood Composite Applications

The aim of this research work was to evaluate the potential of using renewable natural feedstock,i.e.,Jatropha curcas oil(JCO)for the synthesis of non-isocyanate polyurethane(NIPU)resin for wood composite applications.Commercial polyurethane(PU)is synthesized through a polycondensation reaction between isocyanate and poly-ol.However,utilizing toxic and unsustainable isocyanates for obtaining PU could contribute to negative impacts on the environment and human health.Therefore,the development of PU from eco-friendly and sustainable resources without the isocyanate route is required.In this work,tetra-n-butyl ammonium bromide was used as the activator to open the epoxy ring with 3-Aminopropyltriethoxisylane as a catalyst to yield urethane of JCO(UJCO).The UJCO were characterized by Fourier Transform Infra-Red spectroscopy(FTIR)and their oxirane,and hydroxyl values were measured.The result showed that a decrease in oxirane value was found while the hydroxyl value was increased during the time,confirming that the urethane group was formed.The presence of functional groups in FTIR spectra at wave numbers 1732.08,1562.34,and 3348.42 cm^(−1) indicates the functional groups of C=O(urethane carbonyl),–NH,and–OH,respectively confirmed this finding.The potential applications of NIPU in the wood composite were also outlined.

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%.

Some Biological Features of Aphtona whitfieldi Bryant (Coleoptera: Chrysomelidae), an Insect Pest of Jatropha curcas L. in Burkina Faso

Aphthona whitfieldi Bryant (Coleoptera: Chrysomelidae) is a major insect pest of Jatropha curcas L. (Euphorbiacae) in Burkina Faso and other countries in West Africa. The insect pest feeds on the roots and the leaves of the plant. When the attacks are heavy, the plant may lose all its leaves and die off. Unfortunately, little information is available on the biology of this insect pest. A study was conducted on the biology of this insect pest in the Sissili province, South Burkina Faso and resulted in the knowledge of some of the biological features of the insect pest. Aphthona whitfieldi was reared from 13th July to 22th October 2015. Larvae and pupae were collected in J. curcas plantations near Léo, the capital city of the Sissili province, and brought to the laboratory for rearing. The insects were observed daily and the dimensions and the duration of each stage were recorded. We recorded two larval stages (1st and 3rd): a pre-pupa and a pupal stage. The pupa was free and white milk-like. Both the pre-pupa and the pupal stages lasted for five days. The 1st instar larva was smaller than the third one.

Bioactivity and Kinetic Study of <i>Jatropha curcas</i>Essential Oil Extraction Using Supercritical CO₂

This study reports the extraction of Jatropha curcas leaves using supercritical CO2. Experiments were performed varying the pressure (13 and 20 MPa) and the temperature (50°C and 60°C). The model of Sovová for supercritical fluid extraction was fitted to the experimental kinetic extraction curves. Two cell sizes were used and scale up equations compared. GC analysis showed phytol, carvacrol, and hexahydrofarnesyl acetone as major compounds in all the experiments. A maximum yield of 0.95% dry-weight basis was obtained. It was observed a maximum yield (0.95% dry-weight basis) extract obtained at 20 MPa and 50°C. The results indicated that the mass yield increased with the increase of pressure. The bioassays showed that the extract of J. curcas possessed toxicity against Hyalomma lusitanicum.

Isolation of a genomic DNA for Jatropha curcas ribosome inactivating protein and its tobacco transformation

Genomic DNA for Jatropha curcas ribosome inactivating protein （JRIP） was cloned from total DNA of its leaves by polymerase chain reaction （PCR）. The no intron character was confirmed. The plant expression vector pBI121-JRIP was constructed by inserting the JRIP gene into pBI121 plasmid. The recombinant Agrobacterium EHA105 strain harboring pBI121-JRIP was constructed by conducting pBI121-JRIP to strain EHA 105. PCR and Southern blotting were carried out, and the results proved that the JRIP gene was integrated into tobacco genome. It might provide a new material for disease resistance tobacco species breeding.

Insight of Viral Infection of Jatropha Curcas Plant(Future Fuel):A control based mathematical study

Jatropha Curcas Linnaeous(Jatropha Curcas L)is a wonder plant with a variety of applications and enormous economic potential.Biodiesel,an alternative fuel from non edible vegetable oil of Jatropha Curcas plant,has the requisite potential of providing a promising and commercially viable alternative to diesel oil since it has the desirable physicochemical and performance characteristics comparable to diesel.This alternative fuel is eco-friendly,cost effective and has the huge potentiality for the future generations throughout the world.For effective cultivation of this plant, protection from different viral diseases is essential.In this paper,we describe the eco-epidemiological model of the plant Jatropha Curcas L for understanding the disease dynamics which helps to control the viral infection of the Jatropha Curcas plant cell.By this approach,this plant can grow ideally for the renewable green fuel of the future world.