To determine differential expression of genie male sterility A/B lines in Chinese cabbage-pak-choi (Brassica campestris ssp. chinensis Makino var. communis Tsen et Lee), we used the RNA fingerprinting technique, cDNA-...To determine differential expression of genie male sterility A/B lines in Chinese cabbage-pak-choi (Brassica campestris ssp. chinensis Makino var. communis Tsen et Lee), we used the RNA fingerprinting technique, cDNA-AFLP analysis, in different developmental stages and different tissues. While no obvious differential expressions were observed in rosette leaves, florescence leaves, and scapes, some differential expressions were found in alabstrums of A/B lines and among leaves, scapes and alabstrums. We analyzed the al-abstrums collected in different developmental stages with 10 primer combinations. We got a unique band between middle size alabstrums and large alabstrums in B line in one of the ten pair primers, and in another one pair, one band reflecting a higher gene-expression level in A line than that in B line was obtained. No unique bands were found with the other primer combinations. The bands reflecting different gene-expression level were confirmed by Northern hybridization. The results indicated that cDNA-AFLP was a suitable tool for studying differential expression of genie male sterility in plants. SDS-polyacrylamide gel electrophoresis patterns of soluble proteins further verified the difference in A/B lines.展开更多
By means of Agrobacterium-mediated transformation, 43 kanamycin-resistant buds of Chinese cabbage were got. PCR, PCR-Southern blot and dot blot analysis were used to identify and characterize the putative transgenic p...By means of Agrobacterium-mediated transformation, 43 kanamycin-resistant buds of Chinese cabbage were got. PCR, PCR-Southern blot and dot blot analysis were used to identify and characterize the putative transgenic plants. 26 plants had the predicted bands of the fragment of npt Ⅱ gene. Insect bioassays of 4 transformants showed that toxic protein had been translated and the translation levels were different among these transformants.展开更多
A pot experiment and a sandy culture experiment grown with three vegetable crops of Chinese cabbage ( B. chinensis L., cv. Zao\|Shu 5), winter greens ( B.var.rosularis Tsen et Lee, cv. Shang\|Hai\|Qing) and cele...A pot experiment and a sandy culture experiment grown with three vegetable crops of Chinese cabbage ( B. chinensis L., cv. Zao\|Shu 5), winter greens ( B.var.rosularis Tsen et Lee, cv. Shang\|Hai\|Qing) and celery ( A.graveolens L. var. dulce DC., cv. Qing\|Qin) were conducted, respectively. The initial soil and four incubated soils with different extractable Cd (0.15, 0.89, 1.38, 1.84 and 2.30 mg Cd/kg soil) were used for the pot experiment. Five treatments were designed (0, 0.0625, 0.125, 0.250 and 0.500 mg Cd/L) in nutrient solution in the sandy culture experiment. Each treatment in pot and sandy culture experiments was trireplicated. The objectives of the study were to examine Cd accumulation in edible parts of selected vegetable crops, its correlation with Cd concentrations in vegetable garden soil or in nutrient solution, and evaluate the criteria of Cd pollution in vegetable garden soil and in nutrient solution based on the hygienic limit of Cd in vegetables. Cadmium concentrations in edible parts of the three selected vegetable crops were as follows: 0.01—0.15 mg/kg fresh weight for Chinese cabbage, 0.02—0.17 mg/kg fresh weight for winter greens, and 0.02—0.24 mg/kg fresh weight for celery in the pot experiment, and 0.1—0.4 mg/kg fresh weight for Chinese cabbage, 0.1—1.4 mg/kg fresh weight for winter greens, and 0.05—0.5 mg/kg fresh weight for celery in the pot experiment(except no\|Cd treatment). The order of the three test vegetable crops for cadmium accumulation in the edible parts was celery>winter greens>Chinese cabbage in both the pot experiment and the sandy culture experiment. Cadmium accumulation in edible parts or roots of the vegetable crops increased with increasing of cadmium concentration in the medium(soil or nutrient solution). And cadmium concentrations in edible parts of the test vegetable crops were significantly linearly related to the Cd levels in the growth media (soil and nutrient solution). Based on the regression equations established and the limit of cadmium concentration in vegetable products, the thresholds of Cd concentration in the growth medium evaluated was as follows: 0.5 mg/kg soil of extractable Cd for soil and 0.02 mg /L for nutrient solution. The high capacity for cadmium accumulation in the edible parts of different vegetable crops together with the absence of visual symptoms implies a potential danger for humans.展开更多
基金supported by the National Natural Science Foundation of China(39670512)
文摘To determine differential expression of genie male sterility A/B lines in Chinese cabbage-pak-choi (Brassica campestris ssp. chinensis Makino var. communis Tsen et Lee), we used the RNA fingerprinting technique, cDNA-AFLP analysis, in different developmental stages and different tissues. While no obvious differential expressions were observed in rosette leaves, florescence leaves, and scapes, some differential expressions were found in alabstrums of A/B lines and among leaves, scapes and alabstrums. We analyzed the al-abstrums collected in different developmental stages with 10 primer combinations. We got a unique band between middle size alabstrums and large alabstrums in B line in one of the ten pair primers, and in another one pair, one band reflecting a higher gene-expression level in A line than that in B line was obtained. No unique bands were found with the other primer combinations. The bands reflecting different gene-expression level were confirmed by Northern hybridization. The results indicated that cDNA-AFLP was a suitable tool for studying differential expression of genie male sterility in plants. SDS-polyacrylamide gel electrophoresis patterns of soluble proteins further verified the difference in A/B lines.
基金This work was supported by the Ministry of Education and Wuhan Key Sci-Tech Program
文摘By means of Agrobacterium-mediated transformation, 43 kanamycin-resistant buds of Chinese cabbage were got. PCR, PCR-Southern blot and dot blot analysis were used to identify and characterize the putative transgenic plants. 26 plants had the predicted bands of the fragment of npt Ⅱ gene. Insect bioassays of 4 transformants showed that toxic protein had been translated and the translation levels were different among these transformants.
文摘A pot experiment and a sandy culture experiment grown with three vegetable crops of Chinese cabbage ( B. chinensis L., cv. Zao\|Shu 5), winter greens ( B.var.rosularis Tsen et Lee, cv. Shang\|Hai\|Qing) and celery ( A.graveolens L. var. dulce DC., cv. Qing\|Qin) were conducted, respectively. The initial soil and four incubated soils with different extractable Cd (0.15, 0.89, 1.38, 1.84 and 2.30 mg Cd/kg soil) were used for the pot experiment. Five treatments were designed (0, 0.0625, 0.125, 0.250 and 0.500 mg Cd/L) in nutrient solution in the sandy culture experiment. Each treatment in pot and sandy culture experiments was trireplicated. The objectives of the study were to examine Cd accumulation in edible parts of selected vegetable crops, its correlation with Cd concentrations in vegetable garden soil or in nutrient solution, and evaluate the criteria of Cd pollution in vegetable garden soil and in nutrient solution based on the hygienic limit of Cd in vegetables. Cadmium concentrations in edible parts of the three selected vegetable crops were as follows: 0.01—0.15 mg/kg fresh weight for Chinese cabbage, 0.02—0.17 mg/kg fresh weight for winter greens, and 0.02—0.24 mg/kg fresh weight for celery in the pot experiment, and 0.1—0.4 mg/kg fresh weight for Chinese cabbage, 0.1—1.4 mg/kg fresh weight for winter greens, and 0.05—0.5 mg/kg fresh weight for celery in the pot experiment(except no\|Cd treatment). The order of the three test vegetable crops for cadmium accumulation in the edible parts was celery>winter greens>Chinese cabbage in both the pot experiment and the sandy culture experiment. Cadmium accumulation in edible parts or roots of the vegetable crops increased with increasing of cadmium concentration in the medium(soil or nutrient solution). And cadmium concentrations in edible parts of the test vegetable crops were significantly linearly related to the Cd levels in the growth media (soil and nutrient solution). Based on the regression equations established and the limit of cadmium concentration in vegetable products, the thresholds of Cd concentration in the growth medium evaluated was as follows: 0.5 mg/kg soil of extractable Cd for soil and 0.02 mg /L for nutrient solution. The high capacity for cadmium accumulation in the edible parts of different vegetable crops together with the absence of visual symptoms implies a potential danger for humans.
