The Arabidopsis wild type plants transformed with the transporter gene construct (gene + PBI121) has given me the T0 seeds. The T0 seedlings selected with kanamycin (50 mg/L) when placed in soil, grow up to the silique stage after which the plants are getting wilted and eventually die. Anyone could help me with this?
Suhas Karle It could be the transgene per se, or environmental issues (ex. diseases, stresses....). Did you also grow some wild-type side by side with those transgenic plants to see if the environment factors involved?
Dear @Suhas Karle Whether all your T1 plants got wilted at siliquae stage? If so, then there is a greater possibility of management and diseases issues. Even to rule out all these possibilities, you should have grown the parental type as the check! I am attaching a pdf; hope, it could be helpful to you for managing widely known pests and diseases of Arabidopsis.
@Yeun-Yeu Yeu
I tried growing wild type plants too which could grow fine. There is no possibility of environmental issue as I grow the plants in the plant growth chamber under appropriate conditions. Could you suggest any other options to rule out this ?
@Arbind K. Choudhary
Yes, my all T1 plants are getting wilted at silique stage. I will definitely try to check if there is any disease or something. The wild type generation grows well under same conditions.
Suhas Karle
In some cases, transformation of a specific GOI (gene of interest) can be lethal for transgenic lines [1]. Have any other research groups overexpressed this same GOI in Arabidopsis and the transgenic lines are ok?
[1] Article Overexpression of the Novel Arabidopsis Gene At5g02890 Alter...
You can use a mixture of soilrite instead of soil. Once you transplant your plantlets into soilrite, immediately cover them with polythene (make some holes in the polybag too) to maintain humidity. It will help your plants to grow better. Use a half strength hoagland solution for watering every week.
I don't think that a single gene will affect its agronomic performance with regards to damaging whole plants.
Best wishes
Alkesh Hada Yuan-Yeu Yau Arbind K. Choudhary
Thank you all for the help.
We have shown that Kanamycin causes a genome wide methylation of tobacco plants grown in vitro. The promoter region of a gene coding for an auxin binding protein for example gets hyper methylated (inhibition of the expression of the gene). Have a look at our publication In Journal of Biological Chemistry 272, pp 1534-1540, 1997. Frédéric Schmitt et al. Antibiotics Induce genome wide hyper methylation in cultured Nicotiana tabaccum plants.
How did you transferred and sown those seeds? please explain the method, there may be the answer.
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