利用阿拉伯芥 T-DNA Knockout 突變株 , 研究調控葉部形態發育之功能性基因(2/3)
Date Issued
2004-07-31
Date
2004-07-31
Author(s)
DOI
922311B002025
Abstract
By means of a de novo T-DNA tagging vector pPZP202:BAR:SK with high
convenience and efficiency, we have screened many Arabidopsis T1 T-DNA knockout
lines, whose leaf morphology differs obviously from that in wild-type plant. In this
report, we have identified two gain-of-function mutants, which were selected in the T1
generation, crl1-1D (creased leaf 1-1 Dominant) and crl1-2D with a wrinkled curling
inward and narrow leaf phenotype. In addition, the fertility was lower than wild type
and some of the self-crossed progenies ceased to develop further after the first two
leaves were initiated. Using Southern blotting analysis, two T-DNA insertions with
different Southern patterns were detected in each line, indicating that crl1-1D and
crl1-2D were independent T-DNA tagging lines. Inverse PCR (IPCR) identified that
one of the T-DNA insertion sites in each mutant. Noteworthily, the T-DNA insertion
site both are located at the down-stream region of At2g36310 (CRL1), and is
distanced from each other by 307 bp, which encodes a putative inosine-uridine
nucleoside hydrolase (IUNH). Northern blotting analysis using CRL1 coding region as a probe shown that the transcript levels were higher in mutants than that in wild
type. To recapitulate phenotype of the crl1-1D and crl1-2D in wild type, transgenic
plants with CRL1 overexpression were generated. However, no transgenic plant with
wrinkled leaf was observed, instead some 35S::CRL1 seedlings showed phenotype
similar to the loss-of-function mutant wuschel (wus). The WUS transcripts were not
detected by RT-PCR analysis in 35S::CRL1 lines with the wus phenotype, instead a
shoot apical meristem (SAM) growth suppression was found. From these results, we
suggest that overexpression of the CRL1 might affect the SAM activity rather than
leaf morphology.
Subjects
葉部形態
T-DNA 突變株
基因釣取
creasd leaf1-1D
creasd leaf1-2D
Publisher
臺北市：國立臺灣大學植物科學研究所
Type
journal article
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