Cloning and Functional Analysis of SIDP301 Gene in Oryza sativa

水稻是一种重要的粮食作物,也是单子叶植物研究的模式植物,其抗逆相关基因的克隆与功能分析具有重要的理论和现实意义.水稻SIDP301基因编码的蛋白质含有DUF1644家族保守结构域和锌指结构域.实时荧光定量PCR(qRT-PCR)数据表明SIDP301基因在水稻各个组织中均有表达,其中在根和叶中表达水平较高.SIDP301基因的表达受高盐、高温和茉莉酸诱导.与对照相比,超量表达转基因植株的耐盐性不明显,而抑制表达转基因植株对高盐较敏感.进一步用qRT-PCR检测抗逆相关标记基因,结果表明P5CS、DREB2A、Rab16a和Lea3-1在超量表达转基因植株中的表达量高于野生型植株,而在抑制表达转基因植株中这些基因的表达变化不明显.上述结果说明SIDP301是一个抗逆相关基因,其表达量受抑制时会降低水稻的抗逆性.Rice is an important food crop,and it is also a model plant for the study of monocotyledon.It is of important theoretical and practical significance to clone the resistance genes in rice.SIDP301 gene is cloned from rice,which encodes a protein containing a DUF1644 conservative domain and zinc finger domain.Quantitative RT-PCR( qRT-PCR) data showed that SIDP301 was expressed in all tissues and organs of rice and with higher level in roots and leaves.Its expression was induced by stresses including salt,heat and jasmonate.The salt tolerance of SIDP301-overexpressing transgenic plants had no significant difference compared to the wild-type plants.However,SIDP301-suppressing transgenic plants exhibited hyposensitivity to salt stress compared to the wild-type plants.Through expression profiling analysis using qRT-PCR,all of the examined stress-responsive genes( eg. P5 CS,DREB2A,Rab16 a and Lea3-1) were found to be up-regulated in SIDP301-overexpressing transgenic plants compared to the wild-type plants under salt stress,but the levels of gene expression had no significant difference in the SIDP301-suppressing transgenic plants compared to the wild-type plants. These results indicate that SIDP301 is a stress-related gene,and the suppression of its expression was able to decrease the resistance of rice.国家重大科技专项(2014ZX08001-008);; 国家自然科学基金(31560297

Effects of Low Energy Carbon Ion and Nitrogen Ion Implantation on Germinant Rate, Growth Weight and Peroxidase of Stevia rebaudiana Bertoni

以经过碳、氮 (75× 1 0 14 ke V/ cm2 )离子注入处理后的甜叶菊 (Stevia rebaudianaBertoni)种子为材料 ,研究低能离子注入对甜叶菊萌发率、生长量及过氧化物同工酶的影响。结果表明 :(1 )萌发期处理组的萌发率低于对照 ,萌发延迟 ,幼苗成活率不及对照组 ;萌发后期处理组的相对生长量、过氧化物酶活性高于对照组。 (2 )幼苗移至实验地种植后 ,处理组幼苗长势明显优于未处理组 ,处理组 60 d蕾株比明显高于对照组。 (3)过氧化物酶同工酶谱分析发现 ,对照组具有完整 6条基本带的植株数为 1 2 ,具有完整 B1、B2基本带的植株数为 2 0 ;C+处理组分别为 5和 1 1 ,N+处理组分别为 5和 6。统计发现低能离子的注入影响最大的是同工酶谱 B1、B2带。 N+处理组生物学负效应大于 C+处理组The paper reports the effects of low energy Carbon and Nitrogen ion (75×10 14 keV/cm 2 )implantation into S. rebaudiana seeds.on germinant rate, relative growth weight and peroxidase of S. rebaudiana. The results showed: (1)The seed germinant rates were lower and seeding servival rates and deve lopment speeds were slower in the treated groups than those in the control in the initial period of seed germinating;but in the late period of seed germinating the relative growth weight and peroxidase activity were higher in treated groups than those in the control.(2)When seedings were transplanted into experimental field the growth and development were speeded up in treated groups than that in the control.The rate of flower bud plants were higher in treated groups than that in the control. (3)The isozymogram of peroxidase by PAGE showed that there were 12 plants which had 1～6 basic bands and 20 plants which had B1,B2 bands in the control,but only there were 5 plants and 11 plants, respectively, in C + treated groups,and there were 5 plants and 6 plants, respectively, in N + treated groups. The result also showed there were more negative effects by ion implantation into S. rebaudiana in N + treated groups than those in C + treated groups in the periods of seed germinating and plant growth.国家基金委重大项目资助课题!( 1 9890 30 0

Analysis of Differential Proteins Between Bursaphelenchus xylophilus and B. mucronatus Using Two Dimensional Polyacryamide Electrophoresis and Mass Spectrometry

松材线虫是造成松树萎蔫病的病原,对松林威胁很大。其近似种拟松材线虫与松材线虫在形态学上极其相似,却不具致病性。所以,松材线虫和拟松材线虫的快速检测至关重要。应用双向电泳联用质谱技术,研究松材线虫和拟松材线虫的蛋白差异,并对差异蛋白进行MAldI-TOf/MS分析以及数据库鉴定,共鉴定了45个差异蛋白,其中松材线虫22个、拟松材线虫23个。不仅为松材线虫和拟松材线虫的准确鉴定打下基础,差异蛋白的进一步研究可望揭示松材线虫的致病机理。The plane-parasitic nematode Bursaphelenchus xylophilus is the causal agent of pine wilt disease,which is the most serious conifer disease in the world.However,it is difficult to differentiate isolates of B.xylophilus from the closely related species B.mucronatus,which is not pathogenic to pine trees.So it is very important to distinguish B.mucronatus from B.xylophilus rapidly.The author applied the two-dimensional electrophoresis and mass spectrometry to study the proteomics of the two nematodes,and had obtained 45 differentiate proteins,22 of which were from B.xylophilus and others were from B.mucronatus.The results supplied new insights into the rapid identification of B.xylophilus the virulence of B.xylophilus.国家自然科学基金“‘检测管’诱引松材线虫的机理研究”(30470234

Cell biological effects and POD activities of Stevia rebaudianum induced by carbon ion implantation

比较了以能量为100kev,剂量为1015/cm2碳离子注入的甜菊种子萌发过程的细胞学效应和过氧化物酶活性,发现萌发4d的甜菊幼苗核异常细胞率、染色体畸变频率、细胞核、叶绿体、线粒体的损伤皆明显高于对照组,而细胞分裂指数和POD活性显著低于对照组;萌发第10d,上述表征已与对照组呈不显著差异,表明低能碳离子注入甜菊所引起的损伤已基本修复。Through comparing the effects of carbon ion with energy of100kev and dosage of10 15 /cm 2 implatation on Stevia rebaudiana Bertoni germination about cell biological effects and POD activities,we found Stevia rebaudiana Bertoni of4days,whose abnormal cell nucleus,chromosomes,POD activities,the injures of cell nucleus,choroplasts and mitachondria are higher than the control group(CK),then whose division index of Stavia rebaudiana Bertoni seedling cell was lower than that of the control group.When it grew to10days the difference of the above indexes was nitable.It showed that injuries done by low energy carbon ion implatation on Stevia rebaudiana Bertoni has been repaired.国家自然科学基金重大项目资助课题(19890300

Genetic Analysis and Cloning of the Rice Dwarf Mutant d-ss

通过γ射线诱变,从粳稻品种9522的M2代中筛选出一株矮秆水稻(OryzA SATIVA l.)突变体,定名d-SS.d-SS突变体表现为叶色深绿、短宽的叶片、以及小而圆的籽粒.以d-SS突变体与籼稻品种龙特普杂交的f2代群体为基因定位群体,利用IndEl分子标记将d-SS突变位点定位在5号染色体上的IndEl标记zz5-6和zz1343之间,物理距离为412kb.最终通过图位克隆的方法获得了此基因,测序结果表明此基因在编码区发生了两处缺失突变.Rice height character is one of the most important agronomic traits of rice(Oryza sativa L.).Generally,the dwarf varieties with proper plant height have a greater harvest and higher lodging resistance.Identifying new useful dwarf mutant and understanding its regulating mechanism is an important subject for practice rice breeding.In this study,the d-ss,a mutant of Oryza sativa L.spp.japonicacv.9522,was mutagenized by irradiation with 60 Coγ-ray.The d-ss mutant plants display short stem,dark green leaves,compact panicles,and short,round grains.To map the d-ss locus,an F2population generated by crossing between d-ss(japonica)mutant and Longtepu(indica)was analyzed.The d-ss locus was mapped to rice chromosome 5,between the two InDel markers,ZZ5-6and ZZ1343.The region was delimited to about 412kb.At last,the d-ss gene was cloned by map-based cloning.The analysis of sequencing indicated two deletions happened in translated regions of d-ss gene.国家重点基础研究发展计划(973)项目(2012CB126312

A study on transformation of fused gene O_(21)-O_(14)-A_(21)-HBcAg of FMDV epitopes into Medicago sativa

通过农杆菌的介导,以携带O型和A型口蹄疫抗原决定簇融合基因O21-O14-A21-HbCAg的植物表达载体转化苜蓿(MEdICAgO SATIVA)愈伤组织,并再生得到抗性植株。试验建立了苜蓿甘农1号愈伤组织再生系统;优化了农杆菌介导的苜蓿遗传转化系统;获得抗性再生植株4棵;guS报告基因在转化的愈伤组织中得到瞬时表达;抗性植株PCr检测结果证实目的基因已经成功整合到苜蓿基因组中。The fused gene O21-O14-A21-HBcAg that contained both type O and A of FMDV epitopes were introduced into alfalfa(Medicago sativa cv.Gannong No.1)leaf explants via Agrobacterium tumefaciens-mediated transformation.After infection and co-culture resistant plants were obtained from the resistant callus and the genetic transformation and regeneration system of alfalfa were established.The GUS reporter gene was transitorily expressed in transformed callus.52 regenerated individual plants were acquired through Hygromycin screening test.After PCR tests for transgenic plants,four out of seven randomly selected plants were positive.The results of PCR indicated that the O21-O14-A21-HBcAg gene was integrated into the genomes of transformed alfalfa plants.甘肃省教育厅研究生导师基金(0703-11);兰州理工大学博士基金(SB08200602

Expression and Analysis of Mucosal Immunoadjuvant Activity of LTB

通信作者:[email protected][中文文摘]粘膜免疫是预防一些通过吸附粘膜组织侵入机体病原的很好措施,但粘膜免疫通常需要粘膜免疫佐剂,大肠杆菌(Escherichia coli)不耐热肠毒素的B亚基具有很好的粘膜免疫佐剂活性.本研究从大肠杆菌195菌株中扩增出LT(heat-labile enterotoxin)基因,测序后将其B亚基基因与pET32a连接构建了重组质粒pET32a-LTB,SDS-PAGE显示LTB(heat-labile enterotoxin B subunit)在原核细胞中得到表达,Western blotting和人神经节苷脂结合酶联免疫吸附试验(GM1-ELISA)结果表明,重组LTB具有抗原性和GM1结合活性.将其与禽流感M2e表位融合蛋白M2eHBc+混合通过滴鼻途径免疫小鼠,抗体检测结果表明,所表达的LTB能很好地提高共免疫抗原的粘膜和系统免疫应答.[英文文摘]Mucosal immunity is a good measure for preventing pathogenic agents that occur at or emanate from mucosal surfaces invaded the body,but mucosal immunity usually need to mucosal immunoadjuvant.Escherichia coli heat-labile enterotoxin B subunit(LTB) has a good mucosal immunoadjuvant activity.LT gene was amplified by polymerase chain reaction from an enterotoxigenic E.coli 195 strain in this study.Prokaryotic expression vector pET32a-LTB was constructed by LTB subunit gene and pET32a connection after LT gene was sequenced.SDS-PAGE displayed that LTB subunit are expressed in E.coli cells and recombinant LTB inherited the antigenicity and GM1-binding activity of cholerae enterotoxin by Western blotting and GM1-ELISA assay.The mice immunization with mixing of recombinant LTB and avian infuenza virus M2eepitope fusion protein M2eHBc+by intranasal dropping way revealed that LTB can significantly enhance the mucosal and systemic immune responses of mice to co-administered antigen.福建省科技厅重点项目(2009N0051); 福建省自然科学基金项目(2010J01240);; 厦门市科技计划项目(3502Z20103007

Effects of Low-energy Carbon Ion Implantation on the Growth and Development of Chloroplast of Stevia rebaudiana

研究了能量为１００ ｋｅＶ，剂量为１０１５／ｃｍ ２ 的碳离子对甜菊种子萌发率、幼苗生长发育及叶绿体结构的影响．证明被注入种子出现萌发迟缓、生长速度减慢、植株株型变矮和生物量减少等生物学性状变化；幼叶细胞的叶绿体发育分化减慢、基质类囊体形成滞后、基粒数及基粒类囊体片层减少；部分幼叶细胞叶绿体膜破损、基质片层断裂直至叶绿体解体（约占二十分之一）．表明该能量剂量的碳离子注入将影响种子的生长发育，其原因之一是叶绿体发育迟缓和叶绿体的损伤．该研究为低能碳离子对甜菊诱变育种提供参考依据．This paper reports the effects of carbon ion implantation at 100 keV and 10 15 /cm 2 on the seed germination rate, development, growth, and chloroplast structure of Stevia rebaudiana seedling. The result showed the following changes of the biological characters of the ion implanted seeds in:(1) the retardation of the seed germination, the deceleration of the growth speed, the stunt of plant height, and the decrease of biomass;(2)the deceleration of differentiation and growth of the chloroplast in young leaf cells, the retardation in forming thylakoid, the decrease of the grana numbers and thylakoid lamella; and (3) the damage of the chloroplast membrane, the cleavage of stroma lamella and/or the decomposition of chloroplast(ca.5%).The experimental result proved that the carbon ion implantation at the above energy and dose surely influenced the development and growth of S. rebaudiana seed because of the lesion and development retardation of chloroplast. The study suggested a reference to the induced breeding of S. rebaudiana by low energy carbon ion implantation.国家重大项目资助!课题（19890300