CCM: Biolistic genetic transformation of brown cotton shoot tips 09-26-2016

The researchers used the plant shoot tip culture technique and the gene gun-mediated transformation method and the shoot tips of brown cotton Xincaimian5 (a Gossypium hirsutum L. variety cultured in Xinjiang) as materials to establish a biolistic genetic transformation system to genetically transform cotton shoot tips. And they assessed several critical factors: the type of coagulant, the activated charcoal concentration, the kanamycin concentration, the penetration processing time, and the bombardment pressure and distance.



Source: Internet


The results show that each factor can significantly influence the transformation rate of brown cotton shoot tips. The researchers determined an optimal biolistic genetic transformation technology system:

 

    1. Culture shoot tips for 4 h before being bombarded in the osmotic medium

    2. Bombard shoot tips with helium pressure of 7.586 MPa (1100 psi) with a bombardment distance of 9 cm

    3. After culturing for 4 d in the recovery medium, move shoot tips to MSB screening medium containing 8 g·L1agar powder and 125 mg·L1kanamycin, then screened for 25 d to obtain the resistant germs

    4. Add 1 g·L1activated charcoal to the induction medium to induce rooting of the resistant germs to obtain resistant regenerating plants

 

Based on the detections with the PCR and qPCR, the researchers obtained 16 plants which were positive for the transgene from 5200 shoot tips (transformation rate: 0.31%).


The establishment of this technology system lays a foundation for the genetic transformation of brown cotton, and it also provides new materials for naturally-colored cotton breeding.


*Report delivered by Hou Yunpeng, Feng Hongjie, Cheng Wenhan, Zhang Xinyu, Sunjie and Liu Yong Chang from Oasis Eco-Agricultural Key Laboratory of The Xinjiang Production and Construction Crops.




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