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·L－1agar powder
and 125 mg·L－1kanamycin, then screened for 25 d to obtain the resistant germs
4. Add 1 g·L－1activated 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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