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1.
玉米纹枯病胁迫相关miRNA功能研究   总被引:1,自引:0,他引:1  
罗茂  彭华  宋锐  高健  潘光堂  张志明 《作物学报》2013,39(5):837-844
microRNA (miRNA)是一类内源性的、19~24个碱基长度的小分子非编码RNA, 通过碱基互补调控靶基因的表达, 参与调控植物生长发育, 并在多种非生物与生物胁迫响应中发挥重要作用。但目前关于玉米纹枯病胁迫相关miRNA表达调节与功能尚不完全清楚。利用生物信息学挖掘玉米中的microRNA及其靶基因, 筛选到23条新的玉米miRNA, 预测到89个靶基因。本文在前期研究基础上, 以对纹枯病抗性不同的自交系R15和478作为对象, 采用半定量方法对预测获得部分miRNA前体初步鉴定, 采用实时荧光定量PCR手段对R15和478中表达存在差异的成熟miRNA进行抗纹枯病组织特异性表达验证。结果提示, 部分miRNA在玉米抗纹枯病胁迫过程中可能起着重要的调控作用。  相似文献   

2.
利用高通量测序技术鉴定棉纤维发育相关miRNAs及其靶基因   总被引:3,自引:3,他引:0  
miRNA (microRNA)是一类21~24个核酸长度的非编码小分子RNA(sRNA),它主要通过抑制或降解靶基因来调控植物生长发育等过程.试验利用在纤维长度上有显著差异的2个回交自交系BILs (Backcross inbred lines)的0DPA (Days post anthesis)、3 DPA的胚珠和10 DPA的纤维构建6个sRNA文库并进行Solexa 测序.以已公布的棉花D5基因组序列和棉属其他序列为参考,经分析共发现561个miRNAs,其中包括254个已知的miRNAs(属于40个miRNA家族),75个候选的miRNAs和232个新的miRNAs,研究结果极大地丰富了棉属miRNAs.通过miRNA靶基因预测分析发现多数miRNAs负调控其对应的靶基因,少数正调控.KEGG(Kyoto encyclopedia of genes and genomes)注释结果表明miRNA的靶基因在植物激素代谢途径中显著富集.  相似文献   

3.
香蕉冷胁迫相关microRNA差异表达分析   总被引:1,自引:1,他引:0  
研究香蕉miRNA在冷胁迫处理下的动态表达,初步探索miRNA在香蕉冷胁迫过程中的可能作用。利用茎环引物实时荧光RT-PCR (stem-loop qRT-PCR)策略,检测冷胁迫响应miRNA在5℃冷胁迫处理不同时间点香蕉幼苗叶片中的表达模式。结果表明,在检测的15个香蕉miRNAs中,不同的miRNA 在同一冷胁迫时间下的表达量存在很大差异。根据其表达模式,可将15个miRNA分成4组。组1 和组2中的12个miRNA (miR159e, miR159f, miR160-3p, miR397a, miR399j, miR164e, miR444a, miR408, miR5179, miR530, miR535和miR5538),在5&;#176;C冷胁迫处理下表达量总体为上调,其在胁迫过程中的平均表达水平高于对照CK (0 h) ,说明冷胁迫诱导此类miRNA的表达。第3组包括miR156l、miR162a和miR166a,其在5℃冷胁迫处理各时间点的表达量均低于对照,为下调表达,说明冷胁迫抑制此类miRNA的表达。第4组仅含一个miRNA,即miR156a,除在冷胁迫处理12 h表达量明显上调外,其余时间点均下调表达。对这些miRNAs 作用的靶基因进行预测和分析发现,它们主要参与植物的新陈代谢、信号传导和生长发育过程等。这为进一步验证与低温胁迫相关的目的miRNA 的功能提供理论依据。  相似文献   

4.
为筛选炭疽病菌侵染苹果属植物后相关miRNA的响应情况,以抗病品种‘八棱脆’海棠、感病品种‘嘎啦’苹果2个为试验材料,分别用胶胞炭疽孢子悬浮液侵染组培苗,4天后发病,提取植物总RNA并反转miRNA,用qRT-PCR分析2种植物中抗病相关miRNA相对表达量的差异,并对其靶基因进行预测。在抗性品种中miR390a和miR396b/c/f表达量下调,而在感病品种中表达量上调,且其靶基因均有LRR受体样丝氨酸/苏氨酸蛋白,miR482b靶基因为抗性蛋白。miR390a、miR482b和miR396b/c/f可能在苹果炭疽病的侵染过程中起重要作用。  相似文献   

5.
陆地棉种子发育过程中 microRNA 的挖掘与功能研 究   总被引:2,自引:2,他引:0  
 microRNAs (miRNAs)广泛参与调控植物的生长和发育,但这类分子是否对棉花胚珠和纤维的生长发育起到重要的作用,还有待验证。对这些小分子的认识,一般先要从序列信息的获得上开始。应用基因芯片对TM-1 +5 DPA (开花后5 d)胚珠总RNA进行了miRNA 的筛选研究。结果显示,含有352 个探针的芯片成功钓取到199 个陆地棉miRNA,其中绝大多数为首次报道。此外,对miR399 研究发现,它与纤维中磷的含量变化相关; miR169 可能与干旱应激反应有关。  相似文献   

6.
Chilling stress is a major factor limiting the yield of soya bean [Glycine max (L.) Merr.] on a global scale. However, the regulatory network that controls the chilling response of soya bean remains unclear. In the present study, phenotyping and quantitative analyses of miRNAs in soya bean under chilling stress were carried out to determine the impact of environmental constraints on soya bean productivity. Measurements done during soya bean growth in chilling along with the results of field trials indicated that the cultivars Augusta and Fiskeby V responded differently to low temperatures. Although chilling affected the reproductive development of both cultivars, the final seed output remained unchanged. The differential expression of miR169, miR319, miR397 and miR398 under cold stress was detected using ddPCR. Upon chilling in the reproductive stage, we found that these miRNAs had contrasting expression profiles in Augusta and Fiskeby V. A set of candidate target genes was predicted based on degradome sequencing data. A negative correlation was found between the expression of miR169, miR319 and miR398 and their targets in the roots of both cultivars. Our work elucidates the impact of chilling stress on the productivity of two soya bean cultivars and reveals the importance of miRNA involvement in the low temperature response.  相似文献   

7.
小金海棠缺铁胁迫相关miR394a的克隆与表达分析   总被引:2,自引:1,他引:1  
为了研究铁高效植物小金海棠的miRNAs信息及在缺铁处理下miRNA的表达变化,以进一步了解小金海棠缺铁分子调控机制。以改良CTAB法提取的总RNA为模板,从小金海棠中分离得到miR394a。通过Real-time PCR检测miR394a的表达,并利用生物信息学方法预测了miR394a的靶基因及其功能。miR394a具有高度保守性,从小金海棠中分离的miR394a与其他物种的miR394a序列高度相似。缺铁胁迫后,小金海棠的miR394a在根及叶中都有表达,但表达模式有所不同,miR394a在根部响应较为迅速,缺铁处理1天时,即有上调表达;而在叶片的响应较晚,缺铁3天时有上调表达。miR394a的靶基因预测表明,miR394a主要调控转录因子及参与代谢途径的基因。结果表明,miR394a受缺铁胁迫所诱导,参与了小金海棠缺铁调控途径,在缺铁调控中起重要作用。  相似文献   

8.
The use of host resistance (R) genes is considered the most cost‐effective option to control the rice blast disease. The two allelic R genes Pi2 and Pi9 confer very broad‐spectrum resistance against blast isolates collected worldwide. However, the two genes have not yet been widely deployed in rice breeding programmes. Availability of specific markers for them would facilitate incorporating the two R genes into new rice lines through marker‐assisted selection. Herein, we report the development and utilization of a robust and specific marker for the Pi2 and Pi9. This marker was derived from polymorphisms within the target gene, and achieved simultaneously distinguish Pi2 and Pi9 from other alleles through high‐resolution melting of a small amplicon. With the marker, we were able to transfer the Pi2 into an elite restorer line through marker‐assisted backcrossing, successfully obtained effective resistance to blast disease, and we were also able to, respectively, incorporate the Pi2 and Pi9 with two other R genes. As the additive effect, blast resistance in these stacking lines harbouring three R genes were significantly improved.  相似文献   

9.
Two‐line hybrid rice technology is an effective way to increase rice production and improve rice quality. In this study, three bacterial blight (BB) resistance genes, Xa7, Xa21 and Xa23, were introgressed separately into C815S, a popular thermo‐sensitive genic male sterile (TGMS) line to develop five BB‐resistant lines (Hua1005S, Hua1002S, Hua1009S, Hua1006S and Hua1001S) to be resistant against seven races of Xanthomonas oryzae pv. oryzae (Xoo). The two‐line hybrids with heterozygous Xa23 were resistant against seven Xoo strains. But, the hybrids with heterozygous loci for both Xa7 and Xa21 were only resistant against three Xoo strains and were moderately susceptible to the other four strains indicating the role of modifiers influencing the poor expression of dominant BB resistance genes under heterozygous state. Among them, Hua1006S was found to be a promising TGMS line with its higher degree of disease resistance level on account of broad‐spectrum resistance gene Xa23 besides possessing better plant type and rice grain quality features.  相似文献   

10.
Nitrogen (N) deficiency is a major yield‐limiting factor in rice production. The objective of this study was to identify putative QTLs for low‐N stress tolerance of rice, using an advanced backcross population derived from crosses between an indica cultivar ‘93‐11’ and a japonica cultivar ‘Nipponbare’ and genotyped at 250 marker loci. Plant height, maximum root length, root dry weight, shoot dry weight and plant dry weight under two N conditions and their relative traits were used to evaluate low‐N tolerance at the seedling stage. A total of 44 QTLs were identified on chromosomes 1, 2, 3, 4, 5, 6, 8 and 9. Eight intervals on five chromosomes were identified to harbour multiple QTLs, suggesting pleiotropism or multigenic effects according to the contributor of alleles. Some QTL clusters were found in the nearby regions of genes associated with N recycling in rice, indicating that the key N metabolism genes might have effects on the expression of QTLs. Several unique QTLs for relative traits were detected, which suggested the specific genetic basis of relative performance.  相似文献   

11.
用生物信息学挖掘玉米中的microRNAs及其靶基因   总被引:3,自引:1,他引:2  
microRNA (miRNA)是一类内源性的、19~24碱基长度的小分子非编码RNA,通过碱基互补调控靶基因的表达,在多细胞生物的基因表达调控过程中扮演着十分重要的角色。植物中的miRNAs具有高度的保守性,这为通过同源比对发现保守的miRNAs提供了思路和途径。本研究通过对拟南芥、水稻等植物已知的miRNAs与玉米EST和GSS数据库的比对,并设置一系列严格的筛选标准,共筛选到23条新的玉米miRNAs;利用WMD 3在线植物miRNAs靶基因预测软件,对新发现的玉米miRNAs进行靶基因预测,总共预测到89个靶基因,进一步功能分析发现这些靶基因参与玉米的生长发育、信号转导、转录调节、新陈代谢及逆境胁迫响应等调控过程。  相似文献   

12.
In rice, pre‐exposure to sublethal treatment followed by harsh lethal treatment is known to improve tolerance of different abiotic stresses at the vegetative stage within and across generations. Our major aim was to test the phenomenon of thermo‐tolerance at flowering across (trans)‐generations and within generation using rice cultivars contrasting for heat stress tolerance at flowering. To test trans‐generational response, plants were exposed to higher temperature at flowering stage and seeds obtained from previous generations were exposed to heat stress during flowering, which recorded significantly lower fertility when exposed to the same degree of stress in their subsequent generations. A pre‐acclimation to moderately high acclimating temperatures imposed over three different durations during the vegetative and initial reproductive stage showed positive response in the tolerant N22, particularly under severe heat stress (40 °C). This finding indicates the possibility of acquiring ameliorative thermo‐tolerant mechanisms at anthesis, restricted to tolerant genetic backgrounds to combat subsequent harsh conditions within the same generation. However, trans‐generational memory was ineffective in mitigating spikelet sterility losses in both tolerant and susceptible backgrounds. Rice is extremely sensitive to heat stress during flowering; hence, similar exercise across other crops of interest needs to be carried out before generalizing conclusions.  相似文献   

13.
miR164是植物特有的一类miRNA,参与调控植物的生长发育和对逆境胁迫的响应。为深入理解Car-miR164的功能和调控机制,本研究通过全基因组手段鉴定鹰嘴豆Car-miR164家族成员并预测靶基因功能,在PmiREN中获得Car-miR164家族5个成员的序列,通过染色体定位技术表明其位于鹰嘴豆Ca3、Ca4和Ca7三条染色体上。通过多序列比对发现Car-miR164家族5个成员的21 bp成熟序列的一致性很高,仅在5'端第13、17和21个核苷酸处存在差异。二级结构分析表明Car-miR164家族5个成员的前体序列都能形成稳定的茎环结构,成熟的miRNA序列均位于5'端。进化树分析表明水稻、拟南芥、苜蓿、大豆和鹰嘴豆中miR164家族成员主要聚为3个分支,Car-MIR164a与Mtr-MIR164j聚在一个分支,其他4个Car-MIR164聚在另外一个分支。靶基因预测表明Car-miR164家族5个成员的靶基因为鹰嘴豆NAC转录因子家族成员NAC09、NAC11和NAC57。转录组数据分析表明在鹰嘴豆8个不同组织中Car-miR164b/c/e的表达量比Car-miR164a/d高。本实验为进一步研究鹰嘴豆Car-miR164家族成员及其靶基因提供理论依据。  相似文献   

14.
15.
Dwarf mutants are valuable and crucial resources for genetic research and crop breeding programme in rice. In this study, we identified a dwarf mutant derive from tissue culture, which exhibited a delayed heading date and dwarfism under long‐day growth conditions, suggesting the heading date of dwarf mutant was sensitive to day length. Based on 2000 F2 mutant‐like individuals from the cross of the mutant and a Japonica var. ‘IRAT129’, the dwarf gene was finally narrowed into a 512‐kb region near the centromere on chromosome 9. According to the sequence analysis of a delimited region, 21 genes had base alternations either in promoters (15 SNPs) or in coding regions (6 InDels) among 73 annotated genes, and five genes were confirmed sequence alternations resulting from their expression mainly in the vegetative organs. Given to the RNAi plants of the five genes incapable to mimic dwarf and late heading date phenotype, the candidate gene remains to be identified by other genetic or molecular methods. Therefore, all these results give us informative foundation for the day‐length‐sensitive dwarf gene isolation.  相似文献   

16.
Rice (Oryza sativa L.) is one of the most important crops in the world which survives from various abiotic stresses in natural environments with specific stress‐involved genes expressed. Plant sHSPs (small heat‐shock proteins) were reported to respond to abiotic stresses. To improve the understanding of sHSPs in rice, we characterized heat‐shock‐protein gene OsHSP18.6 here. OsHSP18.6 could be induced by diverse stresses, such as drought, salt and cold, especially under heat. The gene was found expressed in root, stem, leaf, internode and spikelet. Overexpression of OsHSP18.6 results in increased thermotolerance and exhibits universal tolerance to stresses tested, including heat, drought, salt and cold. Lower levels of malondialdehyde (MDA) and greater activities of catalase (CAT) and superoxide dismutase (SOD) were observed in OsHSP18.6‐overexpression rice under heat and drought. OsHSP18.6‐overexpression lines indicated decreased sterile rates under hot weather without remarkable changes in most of other agronomic traits compared with wild‐type plants.  相似文献   

17.
We developed 178 recombinant inbred lines from a southern‐by‐spring oat population designated as “TxH.” These lines were genotyped to generate a high‐quality linkage map that resolved 6,902 markers into 21 linkage groups that matched closely with the latest hexaploid oat consensus map. Three major quantitative trait loci (QTLs) affecting heading date were found in locations that are consistent with known QTLs and candidate genes, and two other QTLs affecting heading date were found in novel locations. Five QTLs affecting plant height were found. Both sets of QTLs are responsible for transgressive segregation observed for these two traits. Four QTLs affecting resistance to crown rust, caused by the pathogen Puccinia coronata f. sp. avenae, were identified. Two of these QTLs are consistent with known clusters of rust resistance genes, while two may represent new locations of novel rust resistance genes. A complete set of SNP sequences suitable for generating markers for molecular selection is provided.  相似文献   

18.
毛白杨12种microRNAs的低温胁迫差异表达分析   总被引:2,自引:1,他引:1  
为了探讨microRNAs (miRNAs)在杨树低温胁迫不同时间段的差异表达规律,初步鉴定参与杨树低温胁迫响应的miRNAs,以重要乡土树种毛白杨为材料,进行不同时间段(0、8、14、20 h)的低温胁迫(4℃)处理,分别提取小片段RNAs (<200 nt),利用实时定量PCR技术研究毛白杨在低温胁迫后12种miRNAs的差异表达规律。结果显示,miRNAs的表达在低温胁迫下有显著变化,且呈现动态反应。在低温胁迫下,大部分miRNAs的表达受到抑制。miR168a、miR169ac、miR394a-3p、miR530a的表达量在各个时间段显著下调。该研究初步表明,miRNAs在调控毛白杨对低温胁迫的反应中起着重要作用。  相似文献   

19.
Cold stress is one of the major abiotic factors that influence the productivity and geographical distribution of many agriculturally important crops like Hevea brasiliensis. Cultivation of H. brasiliensis in India is being extended to northeastern regions, where low temperature during winter adversely affects its survival, growth, and productivity. Developing cold-tolerant genotypes is a primary requisite to maximize the productivity under such challenging environmental conditions. However, lack of methods for early evaluation of cold tolerance in the newly developed clones and the extensive time required for assessing their tolerance in the field are major constraints for clonal selection. The present study was initiated with an objective to identify and characterize cold stress responsive miRNAs from H. brasiliensis that show stronger association with cold tolerance. Next generation sequencing using Illumina HiSeq method revealed the expression of 21 and 29 conserved miRNA (from clone RRIM 600) families in cold-stressed and control samples, respectively. Forty-two novel miRNAs were identified from this study. Upon differential expression analysis, eight conserved miRNAs were found commonly expressed in both the samples. When expression analyses were performed subsequently with six selected miRNAs in two Hevea clones (viz. RRII 105 and RRIM 600), miR169 showed a strong association with cold tolerance. miRNAs such as miR482 and miR159 also exhibited association with cold tolerance. This study suggests the possibility of employing these miRNAs as markers for cold tolerance after validation in more number of genotypes with varying levels of cold tolerance.  相似文献   

20.
microRNA(miRNA)是一类内源性非编码小RNA,在植物整个生长发育过程中都起着重要作用,是基因表达的重要调控因子,而microRNA172(miR172)是众多miRNA家族中的重要的一员。为了更直观的认识miR172在植物中所发挥的作用,笔者归纳和总结了miR172在调控植物生长发育过程和响应逆境的研究进展,具体分析了miR172在植物营养生长阶段转变、开花、花器官发育、节间长度、植物商品器官发育以及逆境胁迫响应等诸多过程中发挥的作用。笔者认为miR172在植物中仍有很多未知的功能尚未发掘,因此未来深入挖掘和研究miR172在植物生长过程中更多的作用是一个重要的研究方向,在此基础上深入解析miR172在此过程中的功能、分子调控机制,为人为利用miR172调控植物生长发育奠定理论基础。  相似文献   

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