| Autor: |
TENG Zheng, LI Ming, CUI Yong-zhen, FANG Wei-kuan, LIANG Zhao-xu, HE Shan-shan, LIANG Jun, WU Kai-chao, YE Yan-ping, LI Rong-bai |
| Zdroj: |
Journal of Southern Agriculture; 2014, Vol. 45 Issue 8, p1333-1339, 7p |
| Abstrakt: |
[ Objective ]A cold-regulated gene Cbcor15a isolated from Chorispora bungeana was transferred into sugarcane embryogenic callus by Agrobacterium tumefaciens-mediated method to establish high-efficient genetic transformation system in order to provide basis on breeding sugarcane varieties with cold resistance. [ Method ]Sugarcane variety ROC22 was used as tested material to optimize culture media for callus induction, bud differentiation and plantlet rooting, and screen suitable type and dosage of hormone, PPT and antibiotics dosage, which adapted for exogenous gene transferring to sugarcane. By using a bivalent plant expression vector named pCambia1300-Cbcor15a-bar, Cbcor15a gene was transferred into callus of sugarcane using Agrobacterium tumefaciens-mediated method. [ Result ] The results showed that OD600 0.4 of Agrobacterium tumefaciens and 20 min of infecting time were beneficial to callus differentiation. The optimal dosage of PPT for callus induction and differentiation was 0.50 mg/L. After infected, adding 500.00 mg/L antibiotic Cef in co-culture could effectively inhabit growth of Agrobacterium tumefaciens, while adding 300.00 mg/L Cef could promote callus differentiation, and 200.00 mg/L Cef could promote plantlet rooting. Low dosage of NAA in MS medium was more beneficial to callus differentiation, and KT was superior to 6-BA in promoting cell division. By adding 5.00 mg/L NAA and 70.00 g/L sugar in MS medium could effectively promote plantlets rooting. After the preliminary selection using PPT, 286 anti-PPT regenerated plantlets transferred with Cbcor15a gene were obtained. Eighty-three plantlets with well growing were selected to detect by PCR, out of which four anti-PPT regenerated plantlets were positive. [ Conclusion ]The Agrobacterium tumefaciens-mediated genetic transformation system was established successfully for exogenous gene transferring to sugarcane, and Cbcor15a gene had been integrated into genome of sugarcane. [ABSTRACT FROM AUTHOR] |
| Databáze: |
Complementary Index |
| Externí odkaz: |
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