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1、Analysis of gene expression in anther development ofcytoplasmic male sterile Raphanas sativus L.#51015SHI Shengli, WANG Jianbo*(College of Life Sciences, Wuhan University, Wuhan, 430072, China)Abstract: The anthers in owers perform important functions in sexual reproduction. Seldom studieshave been
2、made to study anther transcriptomes to explore genes controlling anther development. Toanalyze the characteristics of gene expression during the male sterile anther degeneration, we producedtranscriptomes of anthers of radish (Raphanus sativus L.) at three progressive developmental stageswith cDNA-A
3、FLP method. The transcriptomes included at least 2556 unique transcripts, whichinvolved in transcription, signal transduction, DNA synthesis, cell growth and others. The differentialdevelopmental proles of the transcripts signify extensive regulation of gene expression in the antherduring developmen
4、t. The transcriptomes were used to dissect the major abnormality of gene expressionof the sterile anther. First, we categorized and charted the developmental proles of all transcriptspresent in the anther; the transcripts of the sterile anther represent obviously difference contrast to thefertile an
5、ther. Second, we successfully selected genes from hundreds of transcripts encoding proteinsfor transcription, signal transduction, materials transport, and pollen wall formation.Key words: Ranphanus sativus; Cytoplasmic male sterility (CMS); Gene expression200 IntroductionCytoplasmic male sterility
6、(CMS) is a maternally inherited trait characterized by the inabilityof a plant to produce functional pollen. The mutants of male sterile plant represent useful tools tounderstand the molecular and cellular aspects of pollen development. Microarrays and SAGE2530354045analysis revealed that the transc
7、riptome of pollen showed a high proportion of genes that wereenriched or selectively expressed in pollen, with proposed functions in cell wall biosynthesis1.Amagai et al. (2003) used cDNA microarrays for the identification of 90 anther specific genes inArabidopsis and Brassica oleracea. A large numb
8、er of these anther specific genes encodedenzymes involved in cell wall modification and over 20% of the genes had sequence identity toenzymes known to be involved in synthesis or modification of pectin2. However, thegenome-wide information of these specific gene expression patterns in relation to th
9、eirdevelopmental stages is still very limited. No data are available concerning transcriptomecomplexity in developing anther. Analysis of the molecular mechanism controlling thedevelopment and differentiation of reproductive male organs was immensely important sinceknowledge of gene regulation durin
10、g sexual reproduction of the flowering plant may result inpotential applications in agriculture such as hybrid seed production by using male sterility.The radish (Raphanus sativus L.) plants used in this work have the same nuclear genomicbackground. Unlike the abortion of pollen began generally at t
11、he same time in four locules ofCMS anthers in many CMS systems, the development of pollen abortion had two prominentfeatures in the 805 CMS line. First, the four locules of the same sterile anther did not degenerate atthe same time and the type I locule degenerated earlier than type II locules. Seco
12、nd, themicrospores or pollens in the same locule degenerated at different stage3. It suggested thatgenetic defects occurred early in microspore development and affected microspore or pollendevelopment at different stages. The characteristic of the degeneration of 805A pollen abortionreflected the ge
13、netic defects, which were caused by the identical factors in the same cytoplasmicand nuclear genetic background, caused different damage on the different locules development. Inthis paper, to obtain more detailed information on gene expression in anthers, we compared thegene expression profiles of a
14、nthers of the sterile that have distinct defects in pollen development tothose of fertile plants by cDNA-AFLP analysis.50Foundations: The Doctoral Programm of Hiher Education of China (2010014111008).Brief author introduction:SHI Shengli (Birth 1981), Ph.D., focused on plant molecular biology resear
15、chCorrespondance author: WANG Jianbo (Birth 1964), Professer, focused on plant molecular biology -1-1 Materials and methods1.1 Plant materialsThe radish (Raphanus sativus L.) plants were grown in the greenhouse of the Hubei Academyof Agricultural Sciences. The sterile plants (805A) arisen spontaneou
16、sly from the fertile plants55 805B, so they have the same nuclear genome with 805B. They were crossed with normal fertileline 805B, and the trait of male sterility was inherited by all the progeny plants showing therebythat the trait was inherited from the female parent, and has been maintained stably for manybackcross generations. These sterile plants were normal in their phenotype in all aspects except forthat the anthers were reduced in size and did no
