国产伦乱,一曲二曲欧美日韩,AV在线不卡免费在线不卡免费,搞91AV视频

當(dāng)前位置:主頁 > 科技論文 > 基因論文 >

煙草CBL家族基因NsylCBL4和NaylCBL6在非生物脅迫中的功能分析

發(fā)布時(shí)間:2024-07-07 06:28
  鈣離子是植物中的第二信使,它參與調(diào)節(jié)植物的生長(zhǎng)發(fā)育以及生物和非生物脅迫反應(yīng)。而類鈣調(diào)神經(jīng)素B亞基蛋白(CBL)被認(rèn)為是植物特異性鈣信號(hào)傳導(dǎo)途徑中至關(guān)重要的Ca2+傳感器。目前為止,本課題組已從林煙草中鑒定到12個(gè)CBL蛋白。本研究構(gòu)建了林煙草NsylCBL4、NsylCBL6的過表達(dá)材料并進(jìn)一步探究了NsylCBL4和NsylCBL6兩個(gè)基因的功能,主要獲得以下結(jié)果:(1)蛋白序列分析:NsylCBL4的CDS序列編碼234個(gè)氨基酸,其氨基酸序列中具有三個(gè)Ca2+結(jié)合基序的功能性EF手型結(jié)構(gòu)。NsylCBL6的CDS序列編碼211個(gè)氨基酸,其氨基酸序列中具有三個(gè)EF手型結(jié)構(gòu)。另外,NsylCBL6蛋白序列與擬南芥AtCBL7蛋白序列有高度相似性,約71.35%,與楊樹PeCBL3蛋白序列的相似性為72.51%。(2)基因過表達(dá)材料的獲得:通過農(nóng)桿菌介導(dǎo)的葉盤轉(zhuǎn)化法將NsylCBL4轉(zhuǎn)入煙草中,通過半定量PCR篩選出目標(biāo)基因的低表達(dá)和高表達(dá)株系。在24個(gè)株系中,選擇15號(hào)和24號(hào)株系的T2代材料進(jìn)行功能分析。通過浸花法將NsylCBL6基因轉(zhuǎn)入擬南芥中。通過基因組PCR檢測(cè)T1代,通過半...

【文章頁數(shù)】:101 頁

【學(xué)位級(jí)別】:博士

【文章目錄】:
摘要
abstract
Abbreviation
CHAPTER Ⅰ INTRODUCTION
    1.1 STRUCTURE AND MECHANISM OF CBLS
    1.2 FUNCTIONS OF CBLS IN PLANTS
    1.3 MECHANISMS OF THE CBL-CIPK SIGNALLING NETWORK
        1.3.1 Differential calcium response,expression,and localization of CBLs and CIPKs
        1.3.2 Differential interaction and activation of CBL-CIPK complexes
        1.3.3 Differential target activation by specific CBL-CIPK complexes
    1.4 CBLS ARE INVOLVED IN RESPONSE TO BIOTIC AND ABIOTIC STRESS
        1.4.1 Magnesium signalling
        1.4.2 Sodium signalling
        1.4.3 Potassium signalling
        1.4.4 Nitrate signalling
        1.4.5 Phosphorus signalling
    1.5 PROSPECTIVE RESEARCH OF CBL GENES
CHAPTER Ⅱ THE FUNCTIONAL CHARACTERIZATION OF NSYLCBL4 IN ABIOTIC STRESS
    2.1 RESEARCH AIMS AND OBJECTIVES
    2.2 MATERIALS AND METHODS
        2.2.1 Plant materials
        2.2.2 Chemical reagents and kits used
        2.2.3 Plant’s RNA extraction(Hot Phenol method)reagents
        2.2.4 DNA plasmid extraction reagents
        2.2.5 Agrobacterium suspension reagents
        2.2.6 Others reagents
        2.2.7 Antibiotics
        2.2.8 Softwares and databases
        2.2.9 Bioinformatic analysis
        2.2.10 Primers
        2.2.11 Construction of plasmids
        2.2.12 Fragment purification procedures
        2.2.13 Ligation of DNA fragment
        2.2.14 Bacterial strains and vectors used
        2.2.15 Instruments and equipments used
        2.2.16 Plant growth requirements and stress treatments
        2.2.17 Plant genomic DNA extraction(CTAB DNA method)
        2.2.18 Identification of transgenic homozygous lines
        2.2.19 Real-time fluorescence quantitative PCR reaction
        2.2.20 PCR product recovery and purification
        2.2.21 Screening and identification of positive bacterial colony for PCR
        2.2.22 Double enzyme digestion
        2.2.23 Transformation techniques of bacteria
        2.2.24 Transformation in agrobacterium using the freeze-thaw method
        2.2.25 Genetic transformation of tobacco
        2.2.26 Transgenic screening analysis
            2.2.26.1 Actin PCR
        2.2.27 Quantitative real-time PCR(q RT-PCR)setting
        2.2.28 Agrobacterium-mediated transient expression in tobacco
        2.2.29 Plasmid construction for subcellular localization analysis
        2.2.30 Histochemical GUS staining protocol
        2.2.31 Plasmid construction for overexpression vector
        2.2.32 Measurement of physiological parameters
    2.3 RESULTS
        2.3.1 Homological analysis of NsylCBL4
        2.3.2 Identification of35S:NsylCBL4 transgenic tobacco
        2.3.3 Functional analysis of NsylCBL4
        2.3.4 Subcellular localization of NsylCBL4
        2.3.5 NsylCBL4 proteins structure predictions
        2.3.6 Effects of NsylCBL4 on leaf morphology in tobacco
        2.3.7 Osmotic treatments to transgenic NsylCBL4,and its chlorophyll content determination
        2.3.8 Effects of NsylCBL4 on leaf epidermal cell growth and light response curve
    2.4 DISCUSSION
CHAPTER Ⅲ BIOINFORMATICS AND FUNCTIONAL ANALYSIS NSYLCBL6,AND ITS EFFECT ON THE ROOT SYSTEM UNDER LOW NITRATE IN ARABIDOPSIS
    3.1 MATERIALS AND METHODS
        3.1.1 Database search and Sequence analysis
        3.1.2 Homological and structure prediction analysis
        3.1.3 Homologue gene:AtCBL7
        3.1.4 Molecular cloning
        3.1.5 Plasmid constructions and plant transformation
        3.1.6 Constructions of overexpression vectors
        3.1.7 Plant transformation
        3.1.8 Transient expression analysis
        3.1.9 Gene expression analysis
        3.1.10 Preparation of RNA and first-strand cDNA synthesis
        3.1.11 Low NO3
- treatment stress assay
        3.1.12 Chlorophyll content determination by suing the robust method via a microplate reader
        3.1.13 Root morphological analysis and root hair microscopy
    3.2 RESULTS
        3.2.1 Nsyl CBL6 cloning and bioinformatics analysis
        3.2.2 Classification and evolutionary analysis NsylCBL6
        3.2.3 Nsyl CBL6 protein structure and functional analysis
        3.2.4 Co-expression and predicted functional partners
        3.2.5 Overexpression of NsylCBL6 gene from tobacco in transgenic Arabidopsis plants
        3.2.6 Functional characterization of NsylCBL6 in Arabidopsis
        3.2.7 Subcellular localization
            3.2.7.1 Cytoplasm and nucleus location of NsylCBL6 protein
        3.2.8 Effects of nitrate stress on chlorophyll fluorescence parameters and images of transgenic Arabidopsis
        3.2.9 Low nitrate increase root hair density in NsylCBl6 in Arabidopsis
    3.3 DISCUSSION
        3.3.1 Structure and functional analysis
        3.3.2 Subcellular localization of NsylCBL
        3.3.3 Chlorophyll fluorescence
        3.3.4 Effect of NsylCBL6 on the diversity of root system architecture in response to low NO3
-
  • CHAPTER Ⅳ CONCLUSIONS AND PERSPECTIVE
    REFERENCES
    ACKNOWLEDGMENTS
    AUTHOR RESUME



    本文編號(hào):4003294

  • 資料下載
    論文發(fā)表

    本文鏈接:http://lk138.cn/kejilunwen/jiyingongcheng/4003294.html


    Copyright(c)文論論文網(wǎng)All Rights Reserved | 網(wǎng)站地圖 |

    版權(quán)申明:資料由用戶d4573***提供,本站僅收錄摘要或目錄,作者需要?jiǎng)h除請(qǐng)E-mail郵箱bigeng88@qq.com
    女人和大鸡巴视频| 中国美女羞羞黄| 成人网站亚洲二区乱码| 国产麻豆一精品一AV一免费女裸| 97在线最新| 日本最新伦中文字幕| 天天干熟女激情视频 | 日韩欧色一区二区三区| 99国内精品久久久久久久丝袜| 骚逼天天干| 538分类精品视频一区二区| 国产一区二区色呦呦| 日韩一区二区三区不卡视频免费视频| www玖玖大香蕉| 污污污在线看免费国产| 呦呦紧窄免费视频| 无码中文字幕久久婷婷超碰色五月| 亚洲欧美日韩小说动漫成人| 好吊妞17c这里只有精品| 射精网一区二区三区| 老女久久一区二区三区生色av| 日韩精品一区五页视频| 欧美性乱淫| 国产日韩美久久日| 亚洲少妇精品对白一区二区| 久久精品国产久精最新章节| 一二三不卡免费视频网站| 亚洲精品蜜桃| 爆操美女教师网站| 亚欧精品费综合视频网| 开心婷婷五月基地| 欧美动物1区2区3区| 午夜精品大片| 夜夜嗨日AV| 亚洲视频九九大香蕉| 日韩国产推荐一区二区| 欧美一区二区12p| 国产区一区二区三欠久久| 中文字幕无码在线电影| 93久久夜夜精品躁夜躁夜| 超碰caoporn自拍11p|