Cloning and identification of salt inducible genes in arabidopsis thaliana.

• Other Authors: Chan, Yee-kwan.
• Format: Others
• Language: English; Chinese
• Published: 2000
• Subjects: Plants > Effect of salt on; Arabidopsis thaliana > Effect of salt on; Plant gene expression; Plant molecular genetics; Plants > Adaptation; Salinity
• Online Access: http://library.cuhk.edu.hk/record=b5890290; http://repository.lib.cuhk.edu.hk/en/item/cuhk-323264

Chan Yee-kwan. === Thesis (M.Phil.)--Chinese University of Hong Kong, 2000. === Includes bibliographical references (leaves 108-131). === Abstracts in English and Chinese. === Thesis Committee --- p.i === Abstract --- p.ii === Acknowledgments --- p.v === General Abbreviations --- p.vii === Abbreviation for Chemicals --- p.x === Table of Contents --- p.xi === List of Figures --- p.xiv === List of Tables --- p.xv === Chapter 1. --- Literature Review === Chapter 1.1 --- Salinity as a global problem --- p.1 === Chapter 1.2 --- Salinity and agriculture --- p.2 === Chapter 1.3 --- Plant adaptation to salinity --- p.4 === Chapter 1.3.1 --- Salt secretion --- p.6 === Chapter 1.3.2 --- Ion transport --- p.8 === Chapter 1.3.2.1 --- Role of H+-ATPase in salt tolerance --- p.8 === Chapter 1.3.2.2 --- Potassium and sodium uptake --- p.13 === Chapter 1.3.2.3 --- Sodium efflux --- p.15 === Chapter 1.3.3 --- Osmotic adjustment --- p.20 === Chapter 1.3.3.1 --- Accumulation of mannitol --- p.21 === Chapter 1.3.3.2 --- Accumulation of proline --- p.23 === Chapter 1.3.3.3 --- Accumulation of glycinebetaine --- p.23 === Chapter 2. --- Materials and Methods === Chapter 2.1 --- Plant materials and growth conditions --- p.26 === Chapter 2.1.1 --- Surface sterilization of Arabidopsis seeds --- p.26 === Chapter 2.1.2 --- Determination of sub-lethal inhibitory doses of sodium --- p.27 === Chapter 2.1.3 --- Growth conditions of Arabidopsis seeds for total RNA extraction --- p.27 === Chapter 2.1.4 --- NaCl dosage tests --- p.28 === Chapter 2.1.5 --- Expression kinetic tests --- p.28 === Chapter 2.2 --- Isolation of total RNAs --- p.28 === Chapter 2.3 --- Isolation of genes differentially expressed in NaCl concentration by RAP-PCR --- p.30 === Chapter 2.3.1 --- RNA fingerprinting by RAP-PCR --- p.30 === Chapter 2.3.2 --- PCR reamplificatin of RAP products --- p.31 === Chapter 2.3.3 --- Cloning of differentially expressed genes --- p.33 === Chapter 2.3.3.1 --- Ligation of inserts into pCR-Script vector and transformation --- p.33 === Chapter 2.3.3.2 --- Ligation of inserts into pBluescript II KS (+) T-vector and transformation --- p.36 === Chapter 2.3.3.3 --- Screening of recombinant plasmids --- p.37 === Chapter 2.4 --- Sequencing of differentially expressed genes --- p.39 === Chapter 2.4.1 --- DNA cycle sequencing --- p.39 === Chapter 2.5 --- Northern blot hybridization of NaCl inducible genes --- p.40 === Chapter 2.5.1 --- RNA fractionation by formaldehyde gel electrophoresis --- p.40 === Chapter 2.5.2 --- Northern blotting --- p.41 === Chapter 2.5.3 --- Preparation of single-stranded DIG-labeled PCR probes --- p.41 === Chapter 2.5.3.1 --- Isolation of Total RNA --- p.41 === Chapter 2.5.3.2 --- Primer design --- p.42 === Chapter 2.5.3.3 --- PCR amplification of single-stranded DIG PCR probes --- p.43 === Chapter 2.5.4 --- Hybridization --- p.45 === Chapter 2.5.5 --- Stringency washes --- p.46 === Chapter 2.5.6 --- Chemiluminescent detection --- p.46 === Chapter 3. --- Results === Chapter 3.1 --- Determination of sub-lethal inhibitory doses of sodium --- p.48 === Chapter 3.2 --- Isolation of total RNA from A. thaliana treated with sodium chloride --- p.48 === Chapter 3.3 --- Isolation of genes differentially expressed in sodium concentration by RNA arbitrarily primed polymerase chain reaction RAP-PCR --- p.52 === Chapter 3.3.1 --- Differential cDNA fragments identified by RAP-PCR --- p.52 === Chapter 3.3.2 --- PCR reamplification of RAP products --- p.52 === Chapter 3.3.3 --- Cloning of selected RAP-fragments --- p.62 === Chapter 3.4 --- Nucleotide sequence analysis of selected RAP PCR clones --- p.65 === Chapter 3.5 --- Expression pattern analysis of salt inducible genes by northern blot hybridization --- p.75 === Chapter 3.5.1 --- Preparation of single-stranded digoxigenin (DIG)-labeled probes --- p.75 === Chapter 3.5.2 --- Dosage response of NaCl inducible genes --- p.79 === Chapter 3.5.3 --- Expression kinetics of NaCl inducible genes --- p.80 === Chapter 4. --- Discussion === Chapter 4.1 --- Isolation of RAP-PCR targets --- p.93 === Chapter 4.2 --- Expression of NaCl inducible P450 genes --- p.94 === Chapter 4.2.1 --- Cytochrome P450 CYP73A5 --- p.97 === Chapter 4.2.2 --- Cytochrome P450 CYP83A1 --- p.98 === Chapter 4.3 --- NaCl induction gene related to post-transcriptional activities --- p.99 === Chapter 4.3.1 --- Glycine-rich RNA binding protein (BAC F3F19) --- p.100 === Chapter 4.3.2 --- Chloroplast signal recognition particle (54CP) --- p.103 === Chapter 4.4 --- Conclusion --- p.106 === References --- p.108