Cloning Of LeARF8 Gene In Tomato And Analysis It's Spatial And Temporal Expression Character

Tomato(Mill.Syn.Solanum esculentum L.)is ranking the top in the production area and total yield among all vegetable crops in the world and planted widely in China.Auxin plays an important role in plant development and growth.Applying plant development regulators can increase the amount of ovular fibre and improve the ovaries into fruits.ARF(Auxin Respones Factors)is the key factor in auxin signal transcription pathways.It can receive signals transfered by auxin,and activie or repress the downstream genes' expression.Therefore,it is important to study ARF about it's influence on parthenocarpy and improving the character of tomato fruit.In our previous study,we have study about the spatial and temporal expression Character of ARFs genes during tomato fruits' development,meanwhile,we also make series research on the effect of tomato fruits' development and ARFs genes expression character treated by hormones of different types and concentration.The purpose of this study is to provide the helpful tools for understanding moleculer mechanism of the regulation of fruits growth with auxin.The major study results are as following:(1).We have got the total cDNA longth of tomato LeARFS-1 gene in "mini tomato" using PCR technique.The total longth is 2538bp,the sequence was blasted in NCBI and the homology is 94%.It has the typical structure of ARF gene.We have got partial sequence in the ptomoter region of ARF8 gene using TAIL-PCR.(2).Analysising the expression of four ARFs genes(ARF3,ARF4,ARF6 and ARF8)in tomato stem, leave,ovary before blossom,ovary after blossom and different phases in fruit development.The results indicated that ARF8 is expressed in each position and phase,and it had lower content in the ovary after blossom;ARF3 wasn't expressed in leave and ovary.,The four genes were all expressed inâ…¤-â…¦stages during fruit development.(3).HPLC analysis of the content of endogenous hormones in tomato parthenocarpic fruits with the treatment of hormones of different types and concentration.The results indicated that comparing with nomal fruits Put had a higher content in 30 mg·L^-12,4-D treatment and two GA₃ treaments.In 15 mg·L^-12,4-D and 100 mg·L^-1GA₃ treaments,Spd showed a higher content than in CK.The parthenocatpic fruits had highest content of endogenous IAA in 30 mg·L^-12,4-D treatment,and had lower content of endogenous IAA in both GA₃ treatments compared with CK..(5).The expression character of four ARFs genes(ARF3,ARF4,ARF6 and ARF8)in parthenocarpic fruits treated in hormones of different types and concertration measured by RT-PCR.The results showed that the four genes above are all expressed in tomato parthenocarpic fruits.