Construction and Identification of Plasmid pTA—TUB2

1　Introduction　　Benzamidazole fungicide plays an important role in plant disease control.However,as systemic fungi-cide,due to a long time application,it is facing a serious challenge from resistance problem.For instance,the resistance of Cercosporabeticola〔1〕　　　　 ,Penicillium digitatium Sacc.〔2〕Pyricularia oryzaecav〔3〕 to benzami-dazole fungicides was1 0 0 0 ︼g· m L-1.The resistance of wheat scab( Fusarium graminis)〔4〕,Botrytiscinerea〔5〕 and Sclerotinia sclerotiorum〔6…     

Isolation and Characterization of Ell Gene Promoter from Tomato

A fragment of 2000 bp upstream sequence of Ell clone was amplified from genomic DNA of the tomato cultivar Zhongshu- 5. Sequence analysis showed that the upstream contains the regulatory elements： TATA box （-29 - -22）, CAAT box （-193 - -189）, wound, and drought response elements. Expression vectors of Ell promoter gus fusion were constructed with the promoters of 1 200 and 2 000 bp regions, respectively. Transgenic tomato plants were obtained through Agrobacteriummediated transformation. Histochemical analysis of GUS activity in various tissues showed that the two promoters were able to direct fruit-specific gene expression. The expression driven by promoter of 2 000 bp upstream fragment could increase GUS activity with the maturation of tomato fruits. The promoter of -1 200 bp fragment could direct gus gene expression in fruits with the inductions of drought and wounding. The regulatory region for fruit-specificity was probably located in the region of 1 200 bp of 5′-flanking sequence and some positive regulatory elements or enhancers may exist in the region from -1 200 to -2000 bp.     

Transfer and Detection of barstarGene to Maize Inbred Line 18-599 （White） by Particle Bombardment

In China, the purity of maize hybrid strain is discomforting to the development of seed industrialization. Finding a new method for reproduction of maize hybrid strain is necessary. In this study, using particle bombardment, barstar gene was transferred into maize inbred line 18-599 （White）, which is an antiviral and high quality maize inbred line. By molecular detection of the anther of transgenic maize, two plants transferred with barstar gene were gained in this study, which are two restorer lines. The two plants showed normal male spike, and lively microspores. But the capacity of the two restorer lines should be studied in the future. The aim of this study is to find a new method of reproduction of maize hybrid strain using engineering restorer lines and engineering sterility lines by gene engineering technology.     

Preparation, Characterization, and Application of Antiharpin xoo Antibody

Polyclonal antiharpinxoo rabbit antibody has been prepared successfully using purified harpinxoo protein as an immunogen. The ELISA titer of the antiserum against harpinxoo was about 1：2 000. Western blot analysis showed that the antiserum could bind to the expression harpinxoo protein in particular, hrfl, encoding harpinxoo, is an expression in transgenic rice, detected by antiharpinxoo rabbit antibody. The rabbit antibody against harpinxoo can be used to study further about the biological function, harpinxoo localization, and hrfl gene expression in other plants.     

Vision and development in Trichoderma atroviride

Phototropism, the induction of carotenogenesis and reproductive structures, and resetting of the circadian rhythm are controlled by blue light. Trichoderma is used as a photomorphogenetic model due to its ability to conidiate upon exposure to light. In total darkness, T. atroviride grows indefinitely as a mycelium provided that nutrients are not limiting. However, nutrient deprivation and light trigger the conidiation process. A pulse of blue light given to a radially growing colony induc…     

Tagging and Utilization Bruchid Resistance Gene Using PCR Markers in Mungbean

Sixteen mungbean lines were analyzed using 56 random primers. Different DNA bands were detected between Bruchid resistant lines and susceptible lines. According to the cluster results, the 16 lines can be divided into four groups, including brucid resistant wild types, resistant cultivated lines, resistant progenies and a mixed group. BSA method was used to identify DNA markers that related with bruchid resistant gene by using resistant line and susceptible line and their F2 progeny. One codominant marker was identified, which generated a fragment of 1.79 kb in resistant lines and 1.03 kb in susceptible lines. Finally, this codominant marker was considered to be tightly linked with bruchid resistant gene and could be useful in resistant germplasm identification and marker-assisted selection.     

Molecular Comparisons of Marek＇s disease virus strains of different pathotypes for their gⅠ, gE, pp38 and meq genes

Five to ten serotype I Marek‘ s disease virus (MDV1) strains of different pathotypes were compared for their DNA sequences of gⅠ, gE, pp38 and meq genes. The reference strains were vMDV GA and JM, vvMDV RB1B and Mdll(pl6), vv MDV strains 648A and 584A, vaccine strain CVI988/Rispens; Chinese strains were: v MDV strain N, vvMDV strain G2, vaccine strain 814. Only random aa changes were found in 12 positions within gE of 497 aa among 10 analyzed strains and in 10 positions within gⅠ of 355 aa among 5 strains. There was no relationship found between virus pathotypes and aa changes of both glycoproteins. The aa changes happened in 14 positions within meq of 339 aa among 5 compared strains. But a proline deletion at aa # 194 in a proline- rich domain of meq were shared by two vaccine strains CVI988/Rispens and 814, the latter was a non - pathogenic vaccine strain of serotype 1 isolated in 1983 in China. In another hand, vv MDV strains 648A demonstrated 5 unique aa changes and 4 of them also located in the proline - rich region. The pp38 was very conservative among sequenced 10 strains, there were only two positions at # 107 and # 109 with an altered in its 290 aa. All tested MDV1 strains had glufamine at # 107, instead, only vaccine CVI988 had arginine at the position and lost its epitope reactive with Mab H19, to which all ether tested MDV1 strains were positive. However, CVI988 and vMDV GA shared a Mab T65 - recognized epitope when there was glyeine at aa # 109. It was glutamie acid at aa # 109 in all other MDV1 strains which were not reactive with Mab T65. It seems like that there is some relationship between pathotypes and sequences of pp38 and meq, but more strains need to be compared.     

转cry1Ac+CpTI基因水稻对大螟的致死和亚致死效应

 【目的】研究转cry1Ac+CpTI基因水稻对大螟的致死和亚致死效应,为转基因抗虫水稻的生态风险评价提供依据。【方法】采用活体取食生测的方法系统研究苗期、分蘖拔节期、孕穗期和灌浆成熟期4个生育期的转cry1Ac+CpTI基因水稻KF6和ⅡYouKF6对大螟的致死和亚致死效应。【结果】两转基因水稻在苗期对大螟的致死效应最为明显,其对50%和100％个体的致死作用时间最短,大螟在该生育期的转基因水稻上取食3 d及6 d后,校正死亡率最高;其次为分蘖拔节期,两转基因水稻在4.0 d和10.0 d内能分别对50%和100%的大螟个体产生致死作用,在该生育期的两转基因水稻上取食6 d 后,校正死亡率均明显高于孕穗期和灌浆成熟期;KF6在孕穗期对大螟的致死效应显著下降,其对50%大螟个体的致死作用时间显著延长,且取食6 d 后的校正死亡率也显著下降。但ⅡYouKF6对大螟的致死效应并没有明显下降;两转基因水稻在灌浆成熟期对大螟的致死效应显著下降,对50%大螟个体的致死作用时间显著长于其它生育期,取食3 d和6 d后的校正死亡率均显著低于其它生育期;部分大螟能在转基因水稻的孕穗期和灌浆成熟期完成个体发育。不同生育期的两转基因水稻对大螟的亚致死效应也存在显著差异,取食苗期和分蘖拔节期的转基因水稻能抑制大螟幼虫和蛹的发育,降低化蛹率,但对羽化率没有显著影响;取食孕穗期的转基因水稻能显著抑制大螟幼虫的发育,但对蛹历期、化蛹率和羽化率均无影响;灌浆成熟期的转基因水稻对幼虫和蛹的发育速率、化蛹率和羽化率均无明显影响。在任一生育期的转基因水稻上取食,均能降低大螟幼虫的体重和蛹重,但不影响成虫的生殖力。【结论】不同生育期的转基因水稻,对大螟的致死和亚致死效应存在显著差异,随生育期的增加,致死和亚致死效应逐渐减弱。     

转Bt(Cry1Ab)基因对水稻光合特性及光合产物积累的影响

 【目的】明确外源Bt基因插入对水稻倒1叶光合速率及光合作用相关生理特性的影响。【方法】以转Bt基因水稻及亲本水稻为试材,研究盆栽及田间条件下转Bt基因及亲本水稻苗期、拔节期、抽穗期和成熟期倒1叶光合特性及其相关光合酶活性、光合产物积累的动态变化。【结果】转Bt基因及亲本水稻生理活动峰值均出现在拔节期。与亲本水稻相比,转Bt基因水稻苗期叶片净光合速率显著低于亲本;而拔节期和抽穗期,转Bt基因水稻倒1叶净光合速率、叶绿素含量和乙醇酸氧化酶活性显著高于亲本水稻（P＜0.05）,且拔节期转Bt水稻倒1叶气孔导度和胞间CO2浓度也显著高于亲本水稻（P＜0.05）;成熟期,转Bt基因与亲本水稻倒1叶光合特性无显著性差异（P＞0.05）。【结论】与亲本水稻相比,外源Bt基因的插入对水稻叶片光合特性产生短暂影响,但这种影响不具有持续性。     

东北粳稻不同生育期耐冷性分析

研究以140份东北地区粳稻品种(系)为材料,分别对芽期、分蘖期、孕穗期、灌浆成熟期的耐冷性进行鉴定。结果表明,140份材料中,芽期强耐冷性材料有3份,分蘖期耐冷性较好的占20.00%,孕穗期耐冷性较好的占64.28%,成熟期耐冷性较好的占44.42%;对各生育期耐冷性进行相关分析,结果表明分蘖期地上部分生长量与芽期死苗率和孕穗期结实率呈显著或极显著正相关,孕穗期耐冷性与成熟期耐冷性呈极显著正相关。死苗率可作为分蘖期和孕穗期耐冷性鉴定的间接指标;分蘖期耐冷性可作为孕穗期耐冷性的间接指标。     

干旱胁迫对水稻生育性状与生理指标的影响

为了明确水稻不同生育阶段干旱胁迫对生育性状及生理指标的影响,以‘农大3号’（生育期145天）品种为试验材料,利用盆栽的形式,人工控制土壤水势(-75 kPa),分别在水稻不同生育阶段进行了干旱胁迫对水稻生育和生理指标影响的试验研究。结果表明,分蘖期干旱单穴有效穗数减少6.05~9.52个,干旱越早影响越大。无论任何时期干旱胁迫,处理过后均可以导致干物重下降,成熟期下降幅度为11.20~25.94 g/穴,孕穗期>分蘖期>出穗后各时期。叶面积指数的下降则是体现在干旱过后的下一阶段上,随着CK区基部叶片的衰老与黄化,各处理的叶面积指数逐步与CK接近,干旱越早,生育后期差异就越小。干旱对功能叶片（顶部3叶）长度的影响主要体现在分蘖后期、孕穗前期和孕穗中期,这3个阶段干旱,会明显导致顶部3片功能叶片变短。干旱对灌浆期剑叶和倒2叶的光合速率的影响主要体现在分蘖期和孕穗期,孕穗期>分蘖期>出穗期、灌浆期,剑叶光合速率降低幅度为3.25%~26.50%;倒2叶与剑叶的趋势一致,比CK下降3.93%~39.66%,下降幅度大于剑叶,乳熟期、蜡熟期干旱,测定时尚未进行干旱处理,两片叶的光合速率略高于CK。无论任何时期干旱胁迫,干旱过后都会导致叶绿素含量(SPAD)上升。水稻不同生育阶段干旱胁迫,均能导致单穴有效穗数下降、生物产量降低、功能叶片变短、LAI降低、光合速率下降,最终导致产量下降3.57%~50.79%。     

转基因抗虫水稻中BT蛋白表达量的研究(英文)

对转基因抗虫水稻中Bt蛋白表达量进行研究。[方法]应用酶联免疫吸附测定法(ELISA)定量检测转基因抗虫水稻生同生长期不同部位的Bt蛋自表达量。[结果]转基因水稻灌浆期不同组织中Bt蛋白表达绝对含量的高低顺序为:叶片>未成熟种子及颖壳>根>茎杆;在水稻不同的生长发育(分蘖期、抽穗期和灌浆期)阶段,转基因Bt水稻中Bt蛋白的含量有一些变化;一般在水稻生长后期Bt蛋白的浓度有所下降,但幅度不大,对其抗性不会造成太大影响。[结论]该试验对田间害虫的防治以及转基因水稻的育种都具有重要意义。     

转基因抗虫水稻中Bt蛋白表达量的研究

[目的]对转基因抗虫水稻中Bt蛋白表达量进行研究。[方法]应用酶联免疫吸附测定法(ELISA)定量检测转基因抗虫水稻生相同生长时期不同部位的Bt蛋自表达量。[结果]转基因水稻灌浆期不同组织中Bt蛋白表达绝对含量的高低顺序为:叶片>未成熟种子及颖壳>根>茎杆;在水稻不同的生长发育(分蘖期、抽穗期和灌浆期)阶段,转基因Bt水稻中Bt蛋白的含量有一些变化;一般在水稻生长后期Bt蛋白的浓度有所下降,但幅度不大,对其抗性不会造成太大影响。[结论]该试验对田间害虫的防治以及转基因水稻的育种都具有重要意义。     

表达CpTI或CpTI+Bt基因水稻对鳞翅目害虫抗性的田间评价

结果表明,表达CpTI或CpTI+Bt基因水稻可在整个生育期对鳞翅目害虫保持高度或中度抗性.在分蘖期和灌浆期,相应普通水稻已遭受钻蛀性鳞翅目害虫(包括二化螟、三化螟和大螟等)严重危害,枯心率或白穗率均达10%以上;而该转基因水稻在分蘖期和灌浆期的枯心率或白穗率比对照分别下降了69.80%-96.40%和83.40%-93.30%.田间调查表明,相应普通水稻稻叶平均受害指数高达23.52时,转基因水稻稻叶受害指数仅为0.37-4.05,该转基因水稻可有效控制稻纵卷叶螟为害.但该转基因水稻对褐飞虱、电光叶蝉和黑尾叶蝉的抗性与普通水稻未见明显差异.     

不同灌溉模式和施氮处理下稻田氨氧化细菌及无机氮对N2O排放的影响

通过大田试验,研究不同灌溉方式和施氮处理下早稻、晚稻不同时期稻田N_2O的排放通量、氨氧化细菌数量、氨氧化潜势和无机氮含量的变化,揭示土壤氨氧化细菌数量、氨氧化潜势和无机氮含量对稻田N_2O排放的影响。两季试验均设3种灌溉模式,即常规灌溉(CIR)、"薄浅湿晒"(TIR)和干湿交替(DIR),和2种施氮处理,即100%尿素-N(FM1)和50%尿素-N+50%猪粪-N(FM2)。结果表明:晚稻、早稻分蘖期和成熟期土壤氨氧化细菌数量较低,而孕穗期和乳熟期数量较高;晚稻孕穗期、早稻孕穗期和乳熟期土壤氨氧化潜势较高,而晚稻、早稻分蘖期和成熟期土壤氨氧化潜势较低;相同施氮处理下,DIR模式土壤NH_4~+-N含量高于CIR与TIR模式,DIR与TIR模式土壤NO_3~--N含量均显著高于CIR模式;晚稻、早稻孕穗期和乳熟期DIR和TIR模式土壤N_2O排放通量比CIR模式显著提高,FM2处理高于FM1处理;氨氧化细菌和氨氧化潜势与NH_4~+-N含量之间呈极显著正相关关系,N_2O排放通量与氨氧化细菌和氨氧化潜势之间呈极显著正相关关系。因此,N_2O的排放受到氨氧化细菌和氨氧化潜势的直接影响,稻田NH_4~+-N含量大小会影响氨氧化细菌数量和氨氧化潜势,从而间接影响N_2O的排放。     

杂交中稻、再生稻两季增产的施肥技术研究

为提高杂交中稻、再生稻两季单产，多次试验其施肥技术，结果表明，中稻适当减少基、蘖肥用作穗（粒）肥，使其前期促蘖壮蘖，中期稳长，群体适中，后期个体健壮，抗逆力增强，光合效率和N肥利用率均提高，因此头、二季均较“重底早追施肥法”显著增产，尤其与穗型较小的品种、中苗移栽、穴植双株、适当密植和增施N肥、中期增施K肥等措施并用时效果更佳。N素基、蘖、穗肥比例为5：2：3的两季增产率均高于其他参试8种单项增产措施，起关键作用的是穗（粒）肥，随施N总量增加而提高其所占比例，对两季总产的配合效应愈好。穗（粒）肥施期则根据中期长势长相而定，一般宜在孕穗期或齐穗期施用。     

稻曲病侵染机制的初步研究

2009 ～2011年对稻曲病的侵染机制进行了初步研究.结果表明,稻曲病菌厚垣孢子的生存力很强,即使在-20℃的低温条件下也可保持8个月以上的活性,揭示厚垣孢子完全可以成为来年稻曲病的初侵染来源;在水稻的苗期、分蘖期、孕穗期和齐穗期分别对其进行药剂处理,孕穗期处理的防效最高,均超过90％,与其他处理间有极显著的差异,揭示孕穗期是稻曲病菌侵染的关键期;水稻不同播期对稻曲病的发生密切相关,抗性越强的品种分期播种对稻曲病的影响越低.