Primary Study On Sound Effecting The Growth Of Arabidopsis Thaliana In Molecular Biology

The effect of environmental stresses on plant growth is one of important fields focused on scientists. It had been found that environmental stresses could have been a positive or negative influence on plant growth [79]. Now the studies about the effect of sound stimulation on plant in molecule biology are rare compared with the researches on individual, tissue and cell. We primarily studied the differential expressions of arabidopsis thaliana seedlings under sound stimulation by mRNA reverse transcription differential display PCR (DDRT-PCR) with silver staining in this thesis.In this thesis, after arabidopsis thaliana seedlings were stimulated by sound wave, the total RNA was extracted. Then the total RNA as template was reversely transcribed into cDNA. And again, the differential expression gene fragments induced by the sound wave were screened with DDRT-PCR and identified through Northern dot. The primary conclusions of this thesis are as follows:Firstly, arabidopsis thaliana seedlings were stimulated by sound wave with a certain intensity (100 db) and frequency (1000 Hz) for 9 days and each day for 60 min respectively referred to the relative research[28] . The total RNA of the treated group and the control group was extracted by RNase Plant Mini kit. The quality of the total RNA could assure the need of the subsequent tests, and the yield rate of total RNA in the treated group was larger than in the control group. Secondly, through separating these PCR products on polypropylene acyl gel electrophoresis, 6 differentially expressed cDNA fragments were obtained, which were SA3,CA2,SG2-1,SG2-2,SG7-1,SG7-2. Molecular weight of fragments was between 200bp to 600bp respectively. After being re-amplified, the SG2-2band had not outcome, others had the similar molecular weight between on the agarose gel and on the polypropylene acyl gel. SG2-2 was fake positive and discarded.Thirdly, to identify the truth of the differentially expressed cDNA fragments, Northern dot was performed. Among differential fragments acquired, three was found to be positive fragments,SA3, SG7-1 and CA2, whose molecular weight are 270bp,580bp and 370bp respectively. SA3 was differentially expressed and SG7-1 was advantaged expressed, while CA2 was restrained by the sound wave. The positive fragments will be cloned and sequenced, and then analyzed their functions further. The fact that three different types of differentially expressed cDNA fragments had been found indicated that expression of some genes were turned on, meanwhile the stress restrained some genes from expression under this mode of sound stress.Fourthly, with analyzing SA3 and the differential gene fragments SA5-2[65], it was observed that this two cDNA fragments had close molecular weight about 270bp and 290bp and were amplified with the same anchored primer(oligo-d(T)10A) and similar random primer. It was possible that SA3 and SA5-2 have the same base sequences.