Orchard Green Mature-Clover Sweet Salt-resistant Screen, Regerneration And HS Gene Transfer

One third of 2,000,000 acres orchards in Tianjin belong to the salt alkaloid area. Soil Stalinizations, soil fertility decline sand and low productivity of soil result in low fruit output and bad quality. As the fruit tree is easily in the progress of premature senility, the economic efficiency fell lower and lower. So the improvement of the grow condition for soil physics and chemistry nature have be an urgent problem.Many orchards green manure crop has the ability of bearing the salty, and planting green manures could improve the soil characters, such as Agati sesbania, Melilotus suavena and so on. They could enhance the soil fertility; on the other hand, they could also reduce the soil salt content and soil pH.As one kind of fine orchard green manure, M. suavena has a more widespread use. Comparing with clover, M. suavena has higher salt-resistant. Because it belongs to the halophyte, M. suavena which is used to be a kind of fine forage has very strong resistance adversity and the ability of nitrogen fixation.But M. suavena could only bear no more than 0.6% soil salt content and inhabit in the moderate salt alkaloid soil. Moreover M. suavena's rate of seedlings emergence and the rate of nursery stock survival is low. Therefore, enhancing M. suavena salt-resistance to achieve the aim of selective fine breeding is the key for salt alkaloid orchard soil amelioration, especially for moderate and heavy salt alkaloid soil.This research firstly collected 7 different regions M. suavena: Ningxia M. suavena, Chifeng M. suavena, Heilongjiang M. suavena, Xining M. suavena 1, Xining M. suavena 2, Xining M. suavena 3, Tianjin M. suavena. Separately through aseptic culture, salt force disposal and salty tolerance screens, the physiological target after the disposal (cell membrane penetrability, leaf blade drifts away praline content, leaf MDA content, height, fresh weight, dry weight) demonstrate that Tianjin M. suavena, Xining M. suavena 1, Chifeng M. suavena bear salt most strongly. Because Chifeng M. suavena seed germinating rate is high and the variation is small, it is chosen as explants of the gene transfer. Through the selection of different hormones, herbicide and explants, regeneration system gene transferred system was set up and the genetransferrd salt-resistant sweet clove is obtained. Main content as follows: 1. Under the 0.8% salt forces, the cell membrane penetrability of Ningxia M. suavena is the highest. The rest in turn are Xining M. suavena 3＞Xining M. suavena 2＞Heilongjiang M. suavena＞Xining M. suavena l＞Chifeng M. suavena＞Tianjin M. suavena. Under the 0.8% salt forces, the leaf blade drifts away praline content of Xining M. suavena 1 is the highest. The rest in turn are Chifeng M. suavena＞Tianjin M. suavena＞Xining M. suavena 3＞Ningxia M. suavena＞Xining M. suavena 2＞Heilongjiang M. suavena. Under the 0.8% salt forces, the height of Chifeng M. suavena is the highest. The rest in turn are Tianjin M. suavena＞Xining M. suavena l＞Xining M. suavena 2＞Xining M. suavena 3＞Heilongjiang M. suavena＞Ningxia M. suavena. Under the 0.8% salt forces, the height of Tianjin M. suavena is the highest. The rest in turn are Xining M. suavena 1＞Xining M. suavena 3＞Chifeng M. suavena＞Xining M. suavena 2＞Heilongjiang M. suavena. Under the 0.8% salt forces, the fresh weight of Xining M. suavena 1 is the highest. The rest in turn are Chifeng M. suavena＞Tianjin M. suavena＞Xining M. suavena 3＞Ningxia M. suavena＞Xining M. suavena 2＞Heilongjiang M. suavena.Under the 0.8% salt forces, the dry weight of Tianjin M. suavena is the highest. The rest in turn are Xining M. suavena l＞Chifeng M. suavena＞Xining M. suavena 3＞Ningxia M. suavena＞Xining M. suavena 2＞Heilongjiang M. suavena. Under the 0.8% salt forces, the dry weight of Tianjin M. suavena is the highest. The rest in turn are Xining M. suavena l＞Chifeng M. suavena＞Xining M. suavena 3＞Ningxia M. suavena＞Xining M. suavena 2＞Heilongjiang M. suavena.After the measure and screen for the cell membrane penetrability, leaf blade drifts away praline content, leaf MDA content, height, fresh weight, dry weight of the different M. suavena variety, it demonstrates that the rank of salt-resistant is that Tianjin M. suavena, Xining Melilotus Suavenal and Chifeng M. suavena is the 1st level; Xining M. suavena 3 is the 2nd level; Ningxia M. suavena and Xining M. suavena 2 is the 3rd; the Heilongjiang M. suavena is the 4th.All the indexes demonstrate that the salt-resistant of Tianjin M. suavena, Chifeng M. suavena, Xining M. suavena 1 are best of all and could be taken as the primary materials for the gene-transfer.2. The best regeneration system of Chifeng M. suavena: the culture direction for hypocotyls of M. suavena used as explants regeneration was on the medium : MS+4mg / LBA+0.2 mg / LIAA ;The buds of regeneration can be elongated by the medium : MS+1mg / LBA+0.2 mg / LIAA; The seeding growth 2-3cm high was put on the MS medium containing of different concentration. The experimental results suggested that the optimal medium for rooting of adventitious buds is : 1/2MS+0.2 mg / LNAA.As no sprout regenerating the concentration of the herbicide is 2 mg/L, so that concentration is the most ideal screen one.The rate for gene-transfering of M. suavena hypocotyls used as explants (growth point damaged) is 6.35%.3. After the gene-transferring of salt-resistant HS for the Chifeng M. suavena, the aim M. suavena was obtained after the screen of herbicide concentration experiment.