Study On The Physiological And Biochemical Properties, Nutrition And Function Of Omatoes Using Space Mutation Breeding

Tomato contains high amount of vitamin C, lycopene, folic acid and minerals etc., especially the lycopene which attracts more and more consumers for its specific medical value. It is an important "fruit and vegetable" crop of special economic. Demands for tomato going uptrend year by year make it become the second largest vegetable. Space mutation breeding used only in China supplies a favorable chance for breeding new species. It makes use of different environments between upper air and ground to place a premium on changes in biologic germ plasm, which can be the raw material of breeding new species. The objective of this study was to evaluate the physiological and biochemical properties, nutrition and function of tomatoes M1, M2 using space mutation breeding by Chinese SZ-5 spacecraft, and comparisons with their parent CK were done. It can provide basis for large area cultivation, processing, development and utilization of tomatoes even other plants using space mutation breeding, and then studies were promoted further.Part One:Study of differences between tomatoes M1, M2 and CKWe studied methods of genomic DNA of young leaves of tomatoes M1, M2 and CK extraction and different Polymerase Chain Reaction (PCR) amplification systems, and the optimum and improved system of reaction was obtained. DNA sequences were obtained according to the designed primers. The results of sequencing showed that eight base sequences change had taken place between CK and M1, M2. The study of molecular genetics in various materials could directly reflect the effects of space mutation breeding on plant hereditary. Our results demonstrated that there were some differences among CK, M1 and M2 at DNA level.In order to quickly analyze varieties of tomato via space mutation breeding with near infrared spectra. Firstly, principal component analysis was used to analyze the clustering of tomato leaf samples. Then abundant spectral data were compressed by wavelet transform and the model was built with radial basis function neural network. The model regarded the compressed data as the input of neural network input vectors and the training process was speed up. One hundred and five leaf samples of CK, M1 and M2 were selected randomly to build the training model, and forty five samples formed the prediction set. The results showed that, at the wavelength range of 400-1000nm, there were obvious differences in spectrogram among CK, M1 and M2. When in Visible light at 550nm, there were significant differences between M1, M2 and CK; While in the near-infrared part behind 700nm, the reflectance value of M1 and M2 were higher than CK. The discrimination rate of 97.8% was achieved by this method. It demonstrated that there were some differences among CK, M1 and M2 from pointview of spectroscopy.One hundred and five fruit samples of CK, M1 and M2 were built the training model. Characteristics of the pattern were analyzed by partial least square. The model was built with radial basis function neural network and regarded the compressed data as the input of neural network input vectors. Forty five samples formed the prediction set. The results showed that, the reflectance rate in this three kinds of tomatoes has similar trend, while there were obvious differences in spectrogram among CK, M1 and M2 at the wavelength range of 400-1000nm. The reflectance value of CK was lower than M1 and M2. The discrimination rate of these two models was achieved 95.6% and 97.8%. It offered a new approach to the fast discrimination of varieties of tomato via space mutation breeding.Part Two:Evaluation of root exudates assessment on tomatoes using space mutation breedingVarious tomatoes were cultivated using hydroponics method. When in seedling and florescence periods, roots were washed and root exudates were collected, respectively. The active residues were finally purified with XAD-4 resin exchange columns. Low-molecular-weight components in root exudates were analyzed using GC-MS. The results showed there were obvious variations on composition and quantity of root exudates of the same kind of plant but different genotypes, and the same plant at different growing stages. Besides, we also found some pollutants and harmful chemicals among these root exudates. Therefore, more attention should be paid on the safety of tomatoes by space mutation breeding for environment and human.Part Three:Evaluation of antibacterial capacity in vivo of tomatoes using space mutation breedingAntibacterial capacity in vivo of tomatoes were compared according to natural and perforated decay rate. The results showed that, natural decay rate of M2 was lower than CK, and natural anti-trauma and anti-alternata and Botrytis cinerea ability of M2 were better than the CK. While natural decay rate of M1 was higher than CK, and natural anti-trauma and anti-Alternata and Botrytis cinerea ability of M2 were worse than the CK.Activities of some antioxidant enzymes in vivo were determined through various stages of fruit growth, such as SOD, CAT, and MDA. Preliminary study was done to evaluate the mechanism of antibacterial capacity in vivo of tomatoes using space mutation breeding. The results showed that, the SOD and CAT value of M2 was usually higher than that of CK, with minimum value of MDA. The SOD and CAT value of M1 changed larger, but achieved minimum eventually, with maximum value of MDA. These were consistant with the results that M2 had the strongest antibacterial capacity in vivo, while M1 had the weakest ability.Part Four:Evaluation of nutrition and function of tomatoes using space mutation breedingTo determinate the main nutritional components of M1, M2, and CK, HPLC, Atomic absorption spectrophotometer, Automatic amino acid analyzer, UV spectrophotometry and other methods were used to determine the contents of nutriments. The results showed that, compared to CK, there were obvious changes in vitamin C, lycopene, flavonoids, amino acids, pectin, Ca, K and etc.. Especially the content of Vc in tomatoes using space mutation breeding reached 71.72mg/100g, lycopene to 8.99mg/100g, they were nearly the 1.5 times than that of the CK. The tomatoes using space mutation breeding had high nutrition value. Thus, it provided the scientific basis for a great culturing and the further comprehensives utilization.To evaluate the anti-cancer effect of the tomatoes using space mutation breeding, viability of cells were determined by MTT assay, and the cell-cycle profile and apoptosis induction were analyzed by Flow cytometry. The effects of tomatoes on uman colon cancer cell lines, SW480, HT-29, SW620 and human blood disease cell lines, Raji, U937 and K562 were evaluated. The results showed that, Ml and M2 can inhibit cell proliferation of SW480 and HT-29 cells. The maximum inhibitory rate reached 30%, and they can induced these cells cycle arrest. But they were ineffective on SW620 cell, and the inhibitory rate was only about 5%. Besides, M1 and M2 can inhibit cell proliferation of Raji, U937 and K562 cells. The maximum inhibitory rate was up to 16%, and they can induced Raji cell cycle arrest. These data suggested that M2 had the strongest effect on colon cancer cells, while M1 had the strongest effect on blood disease cells, and CK had the weakest effect on both these two kinds of cells.