Lv, Xiaoyi Chen, Liangliang Zhang, Hongyan Mo, Jiaqing Zhong, Furu Lv, Changwu Ma, Ji Jia, ZhenhongĪ fabrication of a novel simple porous silicon polybasic photonic crystal with symmetrical structure has been reported as a nucleic acid biosensor for detecting antifreeze protein gene in insects (Microdera puntipennis dzhungarica), which would be helpful in the development of some new transgenic plants with tolerance of freezing stress. Hybridization assay of insect antifreezing protein gene by novel multilayered porous silicon nucleic acid biosensor. molitor was induced by heat, suggesting that this phenomenon may be common in insects the extent and degree of the influence differ in species that have different living conditions. The expression of insect antifreeze protein genes both in M. Temperature difference of 10 degree C was effective in stimulating Tmafps expression. High temperature (37~42 degree C) for 1h~3h treatments increased Tmafps mRNA level 4.8-fold of the control. molitor were exposed to 5 degree C and 15 degree C for 1h~5h, the mRNA level of Tmafps was over 6-fold and 45-fold of the control at 25 degree C. molitor which is breeding in door with wheat bran all these temperatures stimulated significantly to the expression of Tmafps, while the extent and degree of the temperature stimulation on Tmafps expression were much lower than on Mpafps. punctipennis were exposed to these temperatures for 1h~5h was at least 30-fold of the control at 25 degree C. punctipennis which lives in the wild filed in desert. molitor) and temperature difference (10~30 degree C) all stimulated strongly to the expression of AFP genes (Mpafps) in M. Mild low temperatures (5~15 degree C), high temperature (38~47 degree C for M. The expression of antifreeze protein genes in the two beetles, Microdera punctipennis and Tenebrio molitor that have quite different living environment, under different temperatures were studied by using real-time quantitative PCR. To further detect whether heat also induce the expression of afps in other insects, and to determine the expression profiling of insect afps at different temperatures. However, it is reported that heat can influence the expression of afps in the desert beetle Microdera punctipennis. The expression of AFP genes (afps) is induced by low temperature. HEAT INDUCIBLE EXPRESSION OF ANTIFREEZE PROTEIN GENES FROM THE BEETLES Tenebrio molitor AND Microdera punctipennis.Īntifreeze proteins (AFPs) play important roles in protecting poikilothermic organisms from cold damage. Polymerase chain reaction indicated that the transgenes were integrated in the genome of transformants. The strawberry plants, transformed with both AFP genes, were able to root in MS media containing 50 mg.l-1 Km, while no roots grew from nontransformed plant in this condition.
The antifreeze protein genes, naturally in plasmid pSW1 and modified in plasmid BB, were transformed to strawberry leaf explants by Agrobacterium tumefaciens LBA 4404. The modified gene encoding antifreeze protein isoform HPLC 6 was successfully constructed using codons which were optimally expressed in the strawberry plant. Antibiotics sensitivity test found that shoot regeneration from leaf explant was inhibited more than 90% at the concentration of kanamycin (Km as low as 5 mg.l-1. It was found that the best regeneration condition was MS medium containing N6-Benzyladenine (BA and 2,4-Dichlorophenoxy acetic acid (2,4-D at concentrations of 1 mg.l-1 and 0.2 mg.l-1, respectively. Agrobacterium-mediated transformation of modified antifreeze protein gene in strawberryĭirectory of Open Access Journals (Sweden)įull Text Available The optimum condition for shoot regeneration from leaf explants of strawberry cultivar Tiogar was investigated.