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| 1 | 植物实时荧光定量PCR内参基因的特点及选择显示文摘实时荧光定量PCR(qRT-PCR)具有灵敏度高、特异性强、重复的动态定量范围和高通量等优点,是进行植物基因表达和转录分析最常用的技术手段之一。选择合适的内参基因是正确运用实时荧光定量PCR分析目标基因表达变化的前提。近年来,大量研究表明,内参基因的选择应取决于研究者的实验条件;随着实验条件的变化,内参基因的选择也随之变化。因此,实时荧光定量PCR结果分析的准确性在很大程度上依赖于所选择的内参基因是否适合。该文从内参基因的选择、常用内参基因的特点、新内参基因的挖掘、应用内参基因组合的优点和内参基因的稳定性评价等几方面进行综述,以期为研究者在实验中选择合适的内参基因提供参考和理论依据。 | 袁伟 万红建 杨悦俭 | 2012 | 植物学报2012，47，4： | 99 |
| 2 | 水孔蛋白在细胞延长、盐胁迫和光合作用中的作用(英文)显示文摘水孔蛋白属于一个高度保守的、能够进行跨生物膜水分运输的通道蛋白MIP家族。水孔蛋白作为膜水通道,在控制细胞和组织的水含量中扮演重要角色。本研究的重点是属于PIP亚家族的GhPIP1;2和属于TIP亚家族的γTIP1在植物细胞延长中的作用。使用特异基因探针的Northern杂交和实时荧光PCR技术证明GhPIP1;2和GhγTIP1主要在棉花纤维延长过程中显著表达,且最高表达量在开花后5d。在细胞延长过程中,GhPIP1;2和GhγTIP1表达显著,表明它们在促使水流迅速进入液泡这一过程中扮演重要角色。而且也研究了盐胁迫植物中钙离子对水孔蛋白的影响。分别或一起用NaCl或CaCl2处理原生质体或细胞质膜。结果发现在盐胁迫条件下,水渗透率值在原生质体和质膜颗粒中都下降了,同时PIP1水孔蛋白的含量也下降了,表明NaCl对水孔蛋白的功能和含量有抑制作用。同时也观察了Ca2+的两种不同的作用。感知胁迫的胞质中游离钙离子浓度的增加可能导致水孔蛋白的关闭。而过剩的钙离子将导致水孔蛋白的上游调控。同时实验已经证明大麦的一类水孔蛋白-HvPIP2;1有更高的水和CO2转移率。本研究的目标是确定负责转运水和CO2的关键水孔蛋白的类型,并且提高植物对水的利用率、抵抗胁迫的能力,促进CO2的摄取及吸收、提高植物的产量。 | 杨淑慎 崔丽荣 | 2009 | 生物工程学报2009，25，3： | 13 |
| 3 | 苎麻Actin1基因克隆及其在韧皮部纤维不同发育阶段的表达显示文摘通过cDNA文库的PCR筛选法获得苎麻内源肌动蛋白基因片段,采用苎麻[Boehmeria nivea(L.)Gaud]中苎1号为材料,结合RACE技术获得了苎麻的Actin1蛋白编码基因(BnACTIN1)的cDNA全长序列(GenBank登录号为DQ665832,2009年8月1日公布),该基因全长cDNA序列1782bp,编码区长1134bp,编码377个氨基酸残基。Blastn分析表明,BnACTIN1基因与桑树(Morusalba:DQ785808)、蓖麻子(Ricinus communis:AY360221)、棉花(Gossypium hirsutum:AY305723-305736)等植物的肌动蛋白基因序列有高度的同源性。对推导的氨基酸序列进行分子进化分析表明,该基因与棉花Actin蛋白家族AAP73451.1、AAP73457.1聚为一类,三者之间的亲源关系最近。建立荧光定量PCR体系,用18SrRNA和Histone3内参基因对表达水平均一化,分析BnACTIN1基因在苎麻纤维细胞伸长过程中的表达水平变化。结果表明,在株高97cm时最高,约为株高11、150和220cm时表达量的230倍以上,是株高47cm时期的20倍。本研究揭示的所获得BnACTIN1序列为苎麻的Actin1蛋白编码基因,其最高表达时期开始于纤维快速伸长时期,但稍早于纤维发育的高峰期,推测该基因可能与韧皮部纤维伸长过程中的肌细胞骨架形成有关。 | 马雄风 喻春明 唐守伟 朱爱国 王廷周 朱四元 刘建新 熊和平 | 2010 | 作物学报2010，36，1： | 13 |
| 4 | Actin,干旱胁迫条件下相对可靠的内部参照基因显示文摘很多关于干旱胁迫机制的研究涉及目标基因的差异表达,实时定量PCR技术在探测目标mRNA的变化方面是最为敏感的。为了避免偏差,目标基因的表达常常需要参照一个或多个内标基因进行定量,内标基因一般是在不同的处理条件下表达相对稳定的管家基因,目前使用最多的是actin基因家族,但在不同处理条件下许多actin同源基因表达的相对稳定性并没有经过确认。很多研究表明:使用不合适的管家基因定量目标基因的表达,将对实验结果造成严重偏差,为了阐明该问题和选择最合适的参照基因,检测了在干旱胁迫条件下8个actin种内同源基因的表达稳定性,通过geNorm软件对其稳定性进行分析,发现在所选择的管家基因当中ACT(X16280)1的表达最稳定,而选择不稳定的actin基因作为内标,会严重影响对目标基因相对表达量的估计。因此,在研究干旱胁迫下目标基因的表达水平时,为了实验结果的准确性和可靠性,建议采用ACT(X16280)1或ACT(X16280)2作为内标。 | 张家亮 曹金华 王直华 余灿 刘东华 靳德明 | 2010 | 生物学杂志2010，27，3： | 4 |
| 5 | Analysis of genes differentially expressed during initial cellular dedifferentiation in cotton显示文摘The early phase of phytohormone induction is a vital stage of somatic embryogenesis. This phase includes a key process for acquiring cellular totipotency through cellular dedifferentiation. To unravel the molecular mechanism of cellular dedifferentiation in cotton, we constructed a cDNA library using the suppression subtractive hybridization method. A total of 286 differential cDNA clones were sequenced and identified. Among these clones, 112 unique ESTs were significantly up-regulated during the early phase of phytohormone induction, and 40.2% of the ESTs were first identified. GST was highly expressed from 6 to 24 h after induction with phytohormone treatment. PRPs were predominantly expressed and exhibited distinct expression patterns in different treatments, suggesting that they are closely related to cellular dedifferentiation in cotton. Putative GhSAMS, GhSAMDC, GhSAHH and GhACO3 involvement in SAM metabolism was identified in this library. The analysis of qRT-PCR showed that two remarkable increased expressions of the four SAM-related genes happened during the early phase of phytohormone induction, and that a highly positive correlation existed between GhSAMS and GhSAHH. The highest expression level of GhSAMS might be associated with its reentry into the cell cycle. The histological observations further showed that some cells accomplished cellular dedifferentiation and division within 72 h in 2,4-D treatment, and that cellular dedifferentiation might be regulated through two alterations in SAM-dependent transmethylation activity in cotton. In addition, the expression patterns of differential genes in different treatments disclosed the complicated interaction between 2, 4-D and kinetin. | ZHU HuaGuo TU LiLi JIN ShuangXia XU Li TAN JiaFu DENG FengLin ZHANG XianLong | 2008 | Chinese Science Bulletin2008，53，23： | 3 |
| 6 | 基于基因组和转录组数据的胡椒内参基因鉴定显示文摘为系统筛选适合胡椒基因表达模式分析的内参基因,本研究基于胡椒全基因组和转录组数据,初步筛选出62个候选基因。以胡椒主根、芽尖、叶片、花穗、不同发育时期果实以及接种辣椒疫霉菌的主蔓为材料,利用geNorm、NormFinder和BestKeeper软件系统分析候选基因在不同组织中的表达稳定性,结合目标基因的表达模式验证,鉴定稳定表达最优的内参基因为Myeloid leukemia factor 1(Pn17.1212)。结果表明:该基因是分析胡椒果实发育、辣椒疫霉菌响应相关基因表达模式的理想内参。内参基因的选择不应仅限于持家基因,全基因组和转录组信息可提供更加全面的参考基因数据,内参基因的准确鉴定需综合转录组信息、基因表达稳定性、标记基因的表达模式验证等数据分析的结果,获得的理想内参基因可满足不同研究对表达稳定性的要求。 | 胡丽松 吴思婷 段兴帅 岑怡 苏岳峰 范睿 伍宝朵 郝朝运 | 2020 | 热带作物学报2020，41，10： | 1 |
| 7 | 光质对离体培养条件下绿色棉纤维发育及黄酮合成的影响显示文摘为探讨光质对绿色棉纤维生长发育和黄酮合成的影响,以绿色棉品种绿絮棉1号为材料进行离体培养,比较红光、黄光、蓝紫光、白光和暗处理条件下绿色棉胚珠鲜物质质量、纤维长度、纤维素含量、黄酮含量等生理和分子指标的差异。结果表明:光处理能够促进胚珠鲜物质质量的增加,但抑制了纤维的伸长。在纤维发育前期,各光处理抑制了蔗糖合成酶活性和蔗糖合成酶基因表达;纤维发育后期,白光处理提高了蔗糖合成酶活性,促进了蔗糖合成酶基因表达。红光、黄光和蓝紫光处理显著提高了β-1,3-葡聚糖酶活性(10~20 DAC(Days after culture));红光、黄光和白光均能提高β-1,3-葡聚糖基因表达量,而蓝光则抑制了基因的表达。白光促进了纤维素的合成,而蓝紫光抑制了纤维素的合成。在纤维发育初期(10 DAC),蓝紫光抑制了黄酮合成及其关键酶基因表达;而在发育后期(30~40 DAC),白光处理能够促进黄酮的合成及其关键酶基因表达。可见,光质对绿色棉纤维发育及黄酮合成具有一定的影响。 | 洪亮 张雷 钱森和 孙旭 蔡永萍 林毅 高俊山 郭宁 | 2015 | 棉花学报2015，27，4： | 0 |
| 8 | Genome-Wide Identification of Genes Responsive to ABA and Cold/Salt Stresses in Gossypium hirsutum by Data-Mining and Expression Pattern Analysis显示文摘For making better use of nucleic acid resources of Gossypium hirsutum, a data-mining method was used to identify putative genes responsive to various abiotic stresses in G. hirsutum. Based on the compiled database including genes involved in abiotic stress response in Arabidopsis thaliana and the comprehensive analysis tool of GENEVESTIGATOR v3, 826 genes up-regulated or down-regulated significantly in roots or leaves during salt or cold treatment in Arabidopsis were identified. As compared to these 826 Arabidopsis genes annotated, 38 homologous expressed sequence tags (ESTs) from G. hirsutum were selected randomly and their expression patterns were studied using a quantitative real-time reverse transcription-polymerase chain reaction method. Among these 38 ESTs, about 55% of the genes (21 of 38) were different in response to ABA between cotton and Arabidopsis, whereas >70% of genes had similar responses to cold and salt treatments, and some of them which had not been characterized in Arabidopsis are now being investigated in gene function studies. According to these results, this approach of analyzing ESTs appears effective in large-scale identification of cotton genes involved in abiotic stress and might be adopted to determine gene functions in various biologic processes in cotton. | ZHU Long-fu HE Xin YUAN Dao-jun XU Lian XU Li TU Li-li SHEN Guo-xin ZHANG Hong ZHANG Xian-long | 2011 | Agricultural Sciences in China2011，10，4： | 0 |
| 9 | Suppression Subtractive Hybridization Reveals Different Responses of Two Varieties of Gossypium arboreum L. Under Apolygus lucorum Stress显示文摘Plants reshape their transcriptomes, proteomes and metabolomes in response to insect damage. In this study, we used suppression subtractive hybridization to investigate the transcriptomes of two cotton varieties(CCRI41 and CCRI23) under Apolygus lucorum damage. From the CCRI23 libraries we obtained 92 transcripts and from the CCRI41 libraries we obtained 96 transcripts. 26 and 63 of the transcripts from CCRI23 and CCRI41, respectively, had known functions. Using reverse transcription PCR, we detected expression profile of genes with known functions. Ultimately, we identified eight significantly regulated genes, including one downregulated and four upregulated genes from the CCRI41 libraries, and one downregulated and two upregulated genes from the CCRI23 libraries. Only the gene encoding the polyphenol oxidase(PPO) is involved in plant defense against insect herbivores, and the others are related to improving tolerance to insect damage. Quantitative real-time PCR was used to study changes in expression levels during A. lucorum damage in CCRI23 and CCRI41. Significantly regulated genes from CCRI23 showed a response in CCRI23 but not response in CCRI41. Similarly, significantly regulated genes from CCRI41 showed a response in CCRI41 but not response in CCRI23. The results showed that, among transcriptomes of cotton varieties, there are different responses to A. lucorum damage. | ZHANG Shuai LI Jing Lü Li-min WANG Chun-yi LUO Jun-yu CUI Jin-jie | 2014 | Journal of Integrative Agriculture2014，13，6： | 0 |
| 10 | Genome-wide association study of micronaire using a natural population of representative upland cotton(Gossypium hirsutum L.)显示文摘Background: Micronaire is a comprehensive index reflecting the fineness and maturity of cotton fiber.Micronaire is one of the important internal quality indicators of the cotton fiber and is closely related to the value of the cotton fiber.Understanding the genetic basis of micronaire is required for the genetic improvement of the trait.However,the genetic architecture of micronaire at the genomic level is unclear.The present genome-wide association study(GWAS)aimed to identify the genetic mechanism of the micronaire trait in 83 representa:tive upland cotton lines grown in multiple environments.Results GWAS of micronaire used 83 upland cotton accessions assayed by a Cotton 63 K Illumina Infinium single nucleotide polymorphism(SNP)array.A total of 11 quantitative trait loci(QTLs)for micronaire were detected on 10 chromosomes.These 11 QTLs included 27 identified genes with specific expression patterns.A novel QTL,qFM-A12–1,included 12 significant SNPs,and GhFLA9 was identified as a candidate gene based on haplotype block analysis and on strong and direct linkage disequilibrium between the significantly related SNPs and gene.GhFLA9 was expressed at a high level during secondary wall thickening at 20∼25 days post-anthesis.The expression level of GhFLA9 was significantly higher in the low micronaire line(Msco-12)than that in the high micronaire line(Chuangyou-9).Conclusions: This study provides a genetic reference for genetic improvement of cotton fiber micronaire and a foundation for verification of the functions of GhFLA9. | SONG Jikun PEI Wenfeng MA Jianjiang YANG Shuxian JIA Bing BIAN Yingying XIN Yue WU Luyao ZANG Xinshan QU Yanying ZHANG Jinfa WU Man YU Jiwen | 2021 | Journal of Cotton Research2021，4，2： | 0 |
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