The Study Of Tissue Culture And Its Factors On Chaenomeles Speciosa Nakai

Take the seeds of Chaenomeles speciosa Nakai as plant material; we carried out researches about tissue culture and salt tolerant.1. The most important conclusion about tissue culture, as follows:1.1 Multiply and root of adventitious:Compared KT and TDZ, BA has a better effect on multiply adventitious buds of Chaenomeles speciosa Nakai. The effect of NAA and IBA on multiply adventitious buds is similar. The explants cultured on medium supplemented with BA0.5 mg/l +NAA0.2 mg/l have the highest multiply ability. Reducing the content of MS macro element has a negative effect on multiply adventitious buds. The adventitious buds cultured on medium contained 1/2MS+IBA0.2 mg/l have highest induction root rate, and inducted roots is longer than other combination medium.1.2 Induction and subculture of callus tissue:1.2.1 Callus induction①In the experiment of cotyledons, 2,4-D has a better effect on inducing and subculture callus tissue than that of NAA. A satisfactory result of callus tissue induction and callus tissue growth was achieved on MS medium supplemented with 1mg/l 2,4-D and 0.1mg/l KT.②Callus tissue induction rate of hypocotyls was 87%, and it was higher than that of cotyledons which was 69%. The proportion and the degree of callus vitrifaction would scaled up as increasing the concentration of auxin and BA. Best callus of hypocotyls were obtained on MS medium contained 0.1mg/l 2,4-D and 0.3mg/l BA .③Dark culture incentives callus tissue growth. Longer the days of dark culture, the looser of callus tissue. The colors of cotyledons transferred from dark green to light yellow with increasing dark culture.1.2.2 Subculture of callus tissue①On solid medium, the subculture of cotyledons has different culture methods. If the subculture with long time, callus tissue was cultured on medium supplemented with NAA1.0mg/l+KT0.5mg/l or NAA0.5 mg/l + TDZ 2mg/l, which can obtain better result. On the other hand, callus tissue can growth quickly on medium contained 2,4-D 0.1 mg/l +BA 0.3mg/l and we can get more callus tissue in a short time.②Cultured in liquid medium, callus tissue of cotyledons grew slowly.③Callus tissue of hypocotyls growth most rapidly when cultured on medium contained NAA1mg/l+KT0.3mg/l.④The effect of Vc on reducing browning of callus tissue of Chaenomeles speciosa Nakai was less, but promoted colors of callus tissue turn to green.1.3 Induction adventitious buds Induction adventitious buds from cotyledons, it was essential that added some TDZ and NAA into induction medium. Some adventitious buds were vitrifaction, and the adventitious buds were growth normally when induced from the mediumsupplemented with TDZ 0.5 mg/l + NAA 1.0mg/l.2. The effect of different concentration NaCl on the growth of seeds and callus tissue, the main conclusion as follows:2.1 Callus tissueIn low concentration of NaCl, the growth of callus tissue kept normally level,the content of MDA was lower than check, and the activity of POD and SOD higher than check, the content of soluble sugar and Pro was higher than check. When cultured on a high NaCl concentration, the growth of callus tissue reduced rapidly, the content of MDA was increased, and the activity of POD and SOD reduced; the content of soluble sugar also reduced, but the content of Pro was still increased.2.2 SeedingsWith the increasing of NaCl concentration, the germination of seeds and activity of root system was reduced; except the seeds growth in 0.5‰NaCl, the content of seeds leave's chlorophyll in the others treatment was lower than blank. Firstly the activity of plantlets'SOD and POD increased, subsequently decreased, and highest enzyme activity was obtain when the seeds cultured on medium contained 1‰NaCl. The content of MDA was scaled up with the increasing of NaCl concentration.