AtDEAHb3, a DEAH-box RNA Helicase, Involved in Arabidopsis Female Gametophytic

Development
Author :ZhuDongZi
Tutor:ZhangXianSheng;GaoXinQi
School :Shandong Agricultural University
CLC :Q945.4
TYPE :PhD thesis
Download the PDF Full Text:http://www.topresearch.org/showinfo-178-678041-0.html
Year:2012
Abstract:
Female gametophyte plays a critical role in the sexual reproduction of angiosperms. Successful
development of female gametophytes in higher plants is the consequence of coordination of cell
specification, proliferation, and differentiation. Several mutants and genes that are involved in female
gametophyte development were identified.RNA helicases are involved in mRNA transcription, preRNA splicing, RNA maturation, transportation, translation initiation and ribosomal assembly, and
many other cellular processes, which play an important role in eukaryotic organism development.
Over100RNA helicase genes in Arabidopsis genome were found, but only few of them are being
functional analyzed. Previous studies revealed that RNA helicases were involved in regulating plant
growth, development and stress responses.In our study, two T-DNA insertion lines were identified and
no homozygous insertion was found in their selfed progeny. In the self-pollinated progeny of the
heterozygotic mutant plants, the segregation ratio of heterozygous to wild type plants was about1:1,
instead of the typical3:1. Reciprocal cross analysis revealed that the mutant allei could not be
transmitted through the female gametophyte. There are about50%seeds in the silique of the mutant
compared with the wild type. These results indicate that the mutation has a vital effect on the female
gametophyte development. The T-DNA was inserted in a DEAH-box RNA helicase gene. Arabidopsis
genome codes22DEAH-box RNA helicases. We named this gene as AtDEAHb3.In flower stage12, the
majorities of wild-type embryo sacs are mature and develop to the stage FG6or FG7. However,
about50%female gametophytes from atdeahb3/+plants show delayed development, from FG2to
FG7stages, indicating that the synchrony of female gametophytic development is impaired in
atdeahb3pistils.GUS staining analysis in the pAtDEAHb3::GUS transgenic Arabidopsis plants
revealed that AtDEAHb3express in the various tissues, especially in the apical meristem and ovule.
The expression of AtDEAHb3was detected in the whole process of female gametophytic development
by using RNA in situ hybridization.Arabidopsis GFA1is the core component of U5SnRNP complex
involved in pre-mRNA splicing. It was predicated that GFA1and AtDEAHb3have similar expression
patterns in Arahidopsis development. Furthermore, yeast two-hybrid, BiFC and Pull-Down analysis
revealed that AtDEAHb3could physically interact with GFA1, indicating that AtDEAHb3may play a
role in female gametophyte development by involving in pre-mRNA splicing.In the AtDEAHb3amiRNAi transgenic Arabidopsis lines, the expressions of CYCB genes were down-regulated, which
indicates that AtDEAHb3functions in the mitotic process by regulating CYCB genes expression and
pre-mRNA splicing.