Relationship between differential gene expression patterns in functional leaves of maize (<Emphasis Type="Italic">Zea mays</Emphasis> L.) at milk filling stage and heterosis using cDNA-AFLP

To understand the molecular mechanism of maize heterosis ,differential gene expression patterns in the functional leaves of 35 maize hybrids relative to their parents involving 10 elite inbreds at milk filling stage were analyzed hy using cDNA-AFLP,the correlation analyses of various differential expression patterns with the performance and heterosis of main maize agronomic traits were evaluated.The main results were as follows:For uniparental specific expression ,significant positive correlations were detected with the performance of seed weight per ear and 100-seed weight at 0.01 and 0.05 probability levels respectively.For biparental specific expression ,significant negative correlations were detected with the performance of ear diameter and seed weight per ear at 0.01 probability level .For uniparental specific expression ,significant positive correlations were detected with the heterosis of ear diameter and seed weight per ear at 0.01 and 0.05 probability levls rspectively,For biparental specific expression .significant negtive correlation was detected with theheterosis of ear diameter at 0.05 probability level ,However,for F1-spectific expression,for fragments detected only in one parent and F1,and for fragments detected only in two parents or only in F1,no significant correlation was detected with the performance or heterosis of all agronomic traits surveyed.     

调控根系对玉米幼苗生长及氮素利用的影响

在石英砂培养条件下,研究了切除种子根和阻止不定根下扎,对不同根系构成的玉米形态建成、氮素吸收和利用及吸收根补偿效应的影响。结果表明,玉米地上部形态建成随根系形态建成的抑制而抑制,反映为全株干物重随根的干物重、长度、表面积和体积的减弱而显著下降,根冠比减小。控制不定根扎入砂中但保留种子根（处理Ⅰ）及切除种子根且控制不定根扎入砂中（处理Ⅱ）的植株与对照相比,虽全株氮吸收量显著下降,但通过加强氮素在地上部分配利用及大幅提高氮素吸收效率而体现了较高的氮素利用效率。处理Ⅰ和处理Ⅱ的玉米吸收根表现出强烈补偿生长现象,但不同根系形态性状对吸收根补偿效应的贡献随根系间差异、特别是根的粗细差异而变化。与对照相比,处理Ⅰ植株和处理Ⅱ植株中,补偿效应最强的是根长、其次是根表面积、第三是根体积、根干物质重最弱;但处理Ⅱ与处理Ⅰ比较,根干物质重补偿效应最强、根长和根表面积相近居其次、根体积最弱。表明当玉米根体积差别大时,根长比根干物质重更能体现根系功能,而当玉米根体积相近时,根干物质重比根长对根系功能更有意义。     

Ubi1 intron-mediated enhancement of the expression of Bt cry1Ah gene in transgenic maize (Zea mays L.)

The cry1Ah gene was one of novel insecticidal genes cloned from Bacillus thuringiensis isolate BT8. Two plant expression vectors containing cry1Ah gene were constructed. The first intron of maize ubiqutin1 gene was inserted between the maize Ubiquitin promoter and cry1Ah gene in one of the plant expressing vectors (pUUOAH). The two vectors were introduced into maize immature embryonic calli by microprojectile bombardment, and the reproductively plants were acquired. PCR and Southern blot analysis showed that foreign genes had been integrated into maize genome and inherited to the next generation stably. The ELISA assay to T1 and T2 generation plants showed that the expression of Cry1Ah protein in the construct containing the ubi1 intron (pUUOAH) was 20% higher than that of the intronless construct (pUOAH). Bioassay results showed that the transgenic maize harboring cry1Ah gene had high resistance to the Asian corn borers and the insecticidal activity of the transgenic maize containing the ubi1 intron was higher than that of the intronless construct. These results indicated that the maize ubi1 intron can enhance the expression of the Bt cry1Ah gene in transgenic maize efficiently     

玉米自交系幼胚离体培养及植株再生研究

采用4种不同类型的玉米自交系为供试材料，用MS和AG培养基进行诱导培养。获得胚性愈伤组织，结果表明，基因型和外植体的发育时期对愈伤组织的诱导率有很大影响；不同的2，4-D浓度及山梨醇浓度影响愈伤组织的保持和质量改善；6-BA浓度为2mg／L时，有利于再分化培养，在再分化培养中，获得了再生植株。     

QTL mapping of resistance to sheath blight in maize（Zea mays L.）

Maize sheath blight (Rhizoctonia Solani) is a widely occurring fungus disease with great harm to corn-pro- ducing regions in the world. The first happening of sheath blight in China was reported in Jilin Province as early as in 1966[1]. Since the 1970s, the enlargement of corn- growing regions, the application of maize hybrids, the increasing use of fertilizers, especially the nitrogenous fertilizer, and a higher growth-density, all have caused a quick spread of sheath blight, the occurring …     

The relationship of differential expression of genes in GA biosynthesis and response pathways with heterosis of plant height in a wheat diallel cross

Heterosis in internode elongation and plant height is commonly observed in hybrid plants, but the molecular basis for the increased internode elongation in hybrids is unknown. In this study, midparent heterosis in plant height was determined in a wheat diallel cross involving 16 hybrids and 8 parents, and real-time PCR was used to analyze alterations in gene expression between hybrids and parents. Significant heterosis of plant height and the first internode in length were observed for all 16 hybrids, but the magnitude of heterosis was variable for different cross combinations. Analysis revealed that the heterosis of the first internode was significantly correlated to that of plant height （r = 0.56, P 〈 0.05）, suggesting that the increased elongation of the first internode is the major contributor to the heterosis in plant height. Real-time PCR analysis exhibited that significant difference in heterosis of gene ex- pression was observed among all cross combinations. Moreover, heterosis of the first internode in length was correlated significantly and positively with expression heterosis of KS, GA3ox2-1, GA20ox2, GA20ox1D, GA-MYB and GID1-1, but significantly and negatively with expression heterosis of GAI and GA2ox-1, which is consistent with our recently proposed model of GAs and heterosis in wheat plant height, suggesting the alteration of GA biosynthesis and response pathways might be responsible for the observed heterosis in plant height.     

Gus基因在佛手叶片中的瞬间表达

采用基因枪法对佛手(Citrus medica L.var.sarcodactylis(Noot)Swingle)嫩叶和老叶叶盘进行了Gus基因转化和瞬时表达研究,分析了轰击距离、叶片组织特性对转化效果的影响.结果显示,在7.584 5×106Pa轰击压力下,采用6 cm轰击靶距转化嫩叶可获得比较高的Gus基因瞬间表达频率;老叶的转化效果比嫩叶差;增加轰击次数并不一定能提高瞬间表达频率.     

Study on heterosis of intersubspecies between indica and japonica rice （Oryza sativa L.） using chromosome segment substitution lines

Great heterosis exists in the inter-subspecific crossesbetween indica and japonica rice cultivars[1,2] and the ex-ploitation of this heterosis has long been considered as apromising method to further increase rice yield potential[3].Previous studies indicated that there were large differencesbetween the two subspecies with respect to morphology,isozyme, polymorphism of molecular markers and DNAstructure due to long time genetic differentiation[4,5]. Thosedifferences were identified to be clos…     

玉米1,3,4-三磷酸肌醇5/6 激酶ITPK家族基因的鉴定和分析

 1,3,4-三磷酸肌醇5/6-激酶(ITPK)是一种在动物、植物、线虫中都比较保守的多功能酶, 在生物信号传导及生长发育中起重要作用。为了充分研究玉米中ITPK 家族基因系统分析、全生育期基因表达模式及逆境胁迫表达模式, 利用玉米基因组数据库, 通过生物信息学手段, 鉴定玉米ITPK 家族基因的全序列、定位和编码蛋白, 通过序列比对进行进化和分类分析。利用玉米高通量芯片表达数据进行组织表达差异性、干旱胁迫和病害胁迫表达谱分析。结果表明, 玉米基因组中含有6 个ITPK 家族基因, 分布于玉米的4 条染色体上。MEME 保守结构域分析显示, ZmITPK1-5 均含有3 个保守的ATP-grasp-4 结构域, ZmITPK6 含有两个ATP-grasp-4 保守结构域。进化树分析表明ZmITPK 可分为3 个亚家族。各个发育阶段中, 多数成员在生殖器官或营养器官中均有较高的表达量, 只有ZmITPK1 在所有器官中表达量都不高。ZmITPK2 和ZmITPK3 基因受干旱胁迫处理诱导不同程度上调表达。而在生物胁迫条件下均无显著上调或下调表达。     

Molecular cloning, chromosomal mapping and expression analysis of disease resistance gene homologues in rice (Oryza sativa L.)

Resistance-like sequences have been amplified from first strand cDNA and genomic DNA of rice by PCR using oligonucleotide primers designed from sequence motifs conserved between resistance genes of tobacco andArabidopsis thaliana. 3 PCR clones, designatedOsr1, Osr2 andOsr3 which were 98% identical in nucleotide sequence level, have been found to be significantly homologous to known plant resistance genes and all contained the conserved motifs of NBS-LRR type resistance genes, such as P-loop, kinase2a, kinase3a and transmembrane domain.Southern hybridization revealed that rice resistance gene hornologueswere organized as a cluster in the genome. RFLP mapping using a DH population derived from anindica/japonka cross (Zhaiyeqing 8/Jingxi 17) and an RFLP linkage map assigned two copies ofOsrl and one copy ofOsr3 to the distal position of chromosome 12 where a blast resistance QTL has been mapped previously. Northern blot analysis showed thatOsrl gene was constitutively transcribed in rice leaves, shoots and roots. Further study concerning isolation of full-length cDNAs would be conducive to elucidating the functions of these genes.     

Investigation of gene expression profiles in coronary heart disease and functional analysis of target gene

The research outlined here includes constitution of the differential gene expression profile by means of oligonucleotide gene microarray and functional analysis of the target gene for coronary heart disease (CHD). In a microarray screening experiment, the predominance of inflammation- and immune-related genes is presented in the expression profile of 107 differential genes based on the analysis of gene ontology and gene pathway. IL-8, an inflammatory factor, is identified as one of the genes that were markedly up-regulated in CHD. The plasma level of IL-8 is significantly raised in patients with CHD (n = 30) compared with healthy controls (n = 40), which underscores the clinical relevance of the in vitro finding. The further functional analysis shows that IL-8 affects platelet aggregation percentage, expression of CD62p and platelet aggregation morphology in 12 healthy volunteers to some extent. These findings suggest the relevance of inflammation and immune responses to CHD at the DNA level. Moreover, IL-8 may be involved in the pathogenesis of CHD through the pathway of platelet activation. Supported by the National Natural Science Foundation of China (Grant No. 90409021)     

用基因芯片技术分析干旱胁迫下生殖生长期大麦的基因表达

耐旱性是干旱地区稳定和增加大麦产量的一个关键因素。鉴定出与耐旱性相关的功能基因,一方面可了解大麦的耐旱机理,同时还可以促进利用生物技术来改良大麦的耐旱性。在研究中,2个在耐旱性上具有明显差异的大麦品种Tadmor(耐旱)和WI2291(干旱敏感)被选作材料,采用22000个ESTs(基因表达序列标签)的Affymetrix大麦基因芯片Barley1来分析生殖生长期干旱胁迫下2个大麦材料的差异表达基因。研究结果表明,干旱胁迫下2个大麦材料中有77个共调节基因,其中部分基因已被报道过可能与抗旱性相关。这些基因中已有功能注释的基因按其生物学功能被分为14组,猜测它们是干旱胁迫的响应基因,在抗旱性上可能不起重要作用,或者是必需的但单独不足以提高大麦的抗旱性。进一步比较2个材料差异表达的基因,发现二材料之间有372个受干旱调节基因的差异。这些基因中有功能注释基因的生物学功能中可分为15组,其中一些已被认为与抗旱性相关;而对那些未知功能的基因,推测可能亦在大麦的抗旱性上扮演一定的角色。研究所得结果可为阐述生殖生长期大麦的耐旱性机理提供新的认识。     

Genetic analysis and gene mapping of leafy head （lhd）, a mutant blocking the differen-tiation of rachis branches in rice （Oryza sativa L.）

Flowers, fruits and seeds are products of plant re- productive development and provide the important sources of foods for humans. Therefore, the moleculargenetic mechanisms of floral development have been ahotspot of research of plant developmental biology[1]. Rice is one of the most important staple food crops. Theoutcome of its reproductive development would determine the yield and quality of grains. Rice is also a model plantof cereals. Hence, the study of rice reproductivedevelopment, esp…     

Fine mapping of an incomplete recessive gene for leaf rolling in rice （Oryza sativa L.）

Genetic analysis and fine mapping of genes controlling leaf rolling were conducted using two backcrossed generations （BC4F2, BC4F3） derived from a cross between QMX, a non-rolled leaf cultivar as a recurrent parent, and JZB, a rolled leaf NIL of ZB as a donor parent. Results indicated that leaf rolling was mainly controlled by an incompletely recessive major gene, namely rl（t）, and at the same time, affected by quantitative trait loci （QTLs） and/or the environment. A genetic linkage map was constructed using MAPMAKER/EXP3.0 with eight polymorphic markers on chromosome 2, which were screened by BAS method from 500 SSR markers and 15 newly developed insertion/deletion （InDel） markers. The position of rl（t） was estimated with composite interval mapping （CIM） method using WinQTLcart2.5. Gene rl（t） was mapped between markers InDel 112 and RM3763, and 1.0 cM away from InDel 112 using 241 plants in BC4F2 population. To fine map r（t）, one BC4F3 line with 855 plants was generated from one semi-rolled leaf plant in BC4F2. Four new polymorphic InDel markers were developed, including InDel 112.6 and InDel 113 located between markers InDe1112 and RM3763. Based on the information of recombination offered by 191 rolled leaf plants and 185 non-rolled leaf plants from the BC4F3 line ,we mapped r（t） to a 137-kb region between markers InDel 112.6 and InDel 113. Homologous gene analysis suggested that r（t）was probably related to the process of leaf development regulated by microRNA.     

mRNA差异显示技术研究低温条件下深黄被孢霉基因的表达差异

mRNA差异显示技术已经广泛应用于研究植物、动物和微生物在各种环境条件下基因的差异表达研究.研究以15℃和30℃培养的深黄被孢霉M6-22菌体为材料,利用mRNA差异显示技术研究两种培养条件下基因的表达差异.经实时荧光定量PCR验证,共获得7条差异片段,相似性搜索结果表明它们是6-磷酸葡萄糖异构酶、单糖核苷酸转运蛋白、Ras1鸟苷酸转移因子、依赖于NAD的苹果酸脱氢酶、Δ12-脂肪酸脱氢酶、CLK4关联的丝氨酸/精氨酸丰富蛋白和假定蛋白,涉及糖酵解、蛋白质修饰、信号传导、脂肪酸合成和mRNA加工等生命过程,表明深黄被孢霉M6-22低温适应性是多种途径协同调控的结果.