Vitrification In In Vitro Culture Of Strawberry And Its Recovery Technology Research

In this paper,’Toyonoka’strawberry was taken as test material. Shoot tips of creeping stems were used as explants for tissue culture. The main purpose of the study is to explore the main factors which lead strawberry shoots vitrified in the process of tissue culture, and then to explore the measures of which make the vitrification shoots recovery. The main study content includs (1) by using method of orthogonal design, different combinations of6-BA, agar, ammonium ion (ammonium chloride) were added to subculture medium of shoots of’Toyonoka’strawberry to study their effects on vitrification of strawberry shoots;(2) the physiological and biochemical indexes (tissue water content, relative electrical conductivity, soluble sugar content, chlorophyll content, antioxidant enzyme activity) between vitrification shoots and normal plantlets were determined and compared;(3) by using method of orthogonal design, different combinations of activated carbon, PVA, calcium ion(calcium chloride) were added to subculture medium of the vitrification shoots of’Toyonoka’strawberry to study their effects on strawberry recovery rate;(4) the physiological and biochemical indexes of recovery plantlets were determined and compared with vitrification shoots and normal shoots, in order to verify whether the indexes after treatment reached the level of normal plantlets, and to filter out the most suitable way though which can made the vitrification shoots recovery and then to facilitate the production and application. The main findings of this study are (1) analysis of variance showed that6-BA, ammonium ion, and agar are all have significant impact on strawberry vitrification;6-BA was the most significant impact factor, the second was ammonium ion, and agar affected least; the optimum treatment to reduce vitrification rate effectively was No.2(1g/L6-BA+6g/L agar+1650mg/L ammonium ion) and the vitrification rate noly was26.22%, with higher. proliferation ratio;(2) tissue water content and relative electric conductivity of vitrification shoots were higher than them of normal shoots, soluble sugar on the contrary; chlorophyll content of vitrification shoots was significantly lower than normal shoots, but Ca/Cb in normal seedlings and vitrification shoots were no significant differences; oxide dismutase (SOD) and catalase (CAT) activities of vitrification shoots were reduced compared with.normal seedlings, while peroxidase (POD) activity increased;(3) there were significant differences among9treatments in recovery rate of vitrification shoots; the optimum one was No.9(1g/L activated carbon+2g/L PVA+166mg/L calcium ion) and the recovery rate was89.07%, followed by the4th treatment (0.5g/L activated carbon+166mg/L calcium ion),81.05%, and the8th treatment (1g/L activated carbon+1g/L PVA),73.87%, respectively; analysis of variance showed that calcium ion concentration was the most significant impact factor on strawberry vitrification recovery, the second was calcium ion, and polyvinyl alcohol affected least;(4) significant different physiological and biochemical indexes between the recovery plantlets and vitrification shoots were observed but not between recovery plantlets and normal plantlets, indicating that the level of reversal recovery plantlets had reached the level of nomal shoots’.