| Plant tissue culture and micrografting technology are two ways of tea plant propagation.Tea plant tissue culture is prone to pollution,Browning and difficult to take root.Traditional grafting is subject to seasonal temperature limitation,micrografting can combine the two and solve the above problems at the same time.Scion was obtained by tissue culture and rootstock was obtained by cutting root,which could make full use of the characteristics of tissue culture,rapid propagation,genetic stability and strong resistance of rootstock.This study for spring varieties‘plateau green’material,to explore the basic conditions for the transformation and establish’plateau green tea tree stem section for earth quickly get no root robust seedlings as explants technology system,to explore the factors influencing the micro grafts take tea tree,as well as the effects on enzyme activity and amino acids,provide support for superior varieties of tea tree grafting factory nursery,genetic transformation to lay the foundation for tea tree,It provides reference for the study of physiological and biochemical effects of micrografting on tea plant.The main results are as follows:1.Taking’Plateau Green’stem segment with axillary buds as explants,explore the effects of sampling period,disinfection method and plant growth regulator on axillary bud germination,proliferation and seedling strength of explants.The results were as follows:after soaking the stem segment in 75%alcohol for 60s and 0.1%hg for 8min,the survival rate,contamination rate and Browning rate of explants were 91.11%,4.44%and 4.44%respectively after 28d.The survival rate was the highest at the end of March,and 88.67%at 5 weeks.The optimal medium for axillary bud germination was MS+2mg/L 6-BA+1mg/L GA3,and the germination rate was 81.11%after 40 days,the average bud length was 1.93cm,and the seedlings grew well.The optimal medium for proliferation culture was MS+4mg/L 6-BA+2m/L IBA,50d increment coefficient 3.37,seedling strong.The optimal medium was MS+1mg/L 6-ba+0.5m/L IBA,and the seedling height increased by 2.25cm after 50 days.2.The leaves and stem segments without axillary buds of’Plateau Green’were used as explants to explore the effects of different explants types,different disinfection time and plant growth regulators on their survival,callus induction and adventitious bud induction.The results showed that the survival rate,pollution rate and browning rate of leaves and stems soaked in 75%alcohol for 30s and 0.1%hg for 6min were86.67%,9.33%and 4.67%respectively.Then,stem and leaf were inoculated in callus induction medium with different regulator ratio.Under MS+0.5mg/L 6-BA+2mg/L 2,4-D or MS+1mg/L 2,4-D,both stem and leaf could get 100%callus after 30 days.After one subculture,the calli were transferred to the medium for adventitious bud induction,and 13.33%of the calli were induced by MS+3mg/L 6-BA+2mg/L NAA for 60d,but no bud induction was found in the leaves.3.Obtain rootless seedlings with the"Plateau Green"tissue culture as the stock,and cutting raw seedlings’Qianmei 601’as the rootstock,conduct micro-grafting,select the split method,cultivate under 25°C conditions.Before grafting Shoot 2d.Study the scion-banded leaf number,scion top,scion’s lignification degree,group cultivation of seedling seedling time,growth adjustment material concentration,scion length,group cultivation of seedling stem poles and rootstock’s seedling stem poles,etc.Influence.The results showed that the scion with three leaves was easy to survive grafting.The best training time for tissue culture seedlings was 4d.The scion with the top left germinated faster after grafting.The grafting of lignified scion can avoid dying;The scion length was 2.5±0.1cm,and the grafting survival rate was the highest after soaking the scion in 1mg/L NAA for 5min.The rootstock and scion with similar diameter of stem were selected for micrografting and the survival rate was high.Under this study conditions,the maximum survival rate of tea trees is 68.33%.4.After micro-grafting into some physiological biochemical measurements,compared with unlocated seedlings,the overall trend of SOD,CAT,and PAL is the same.The SOD of 21d is 104.28 U·mgprot-1,CAT is 686.00 U·gprot-1,PAL is37.51U·gprot-1;Detecting amino acids of micro-grafted,ungrafted rootstock and ungrafted scion.The results showed that a total of 16 kinds of free amino acids were detected in micrografted,with a total of 1201.95 mg/kg,among which theanine 473.56mg/kg.12 kinds of free amino acids were detected in ungrafted stock’Qianmei 601’,with a total of 369.67 mg/kg,including theanine 36.62 mg/kg.A total of 15 amino acids were detected in ungrafted scion’Plateau green’,with a total of 1267.54 mg/kg,among which theanine 356.54 mg/kg.Micro-grafting seedlings are higher than the unfracorbed rootstock and scion,but phenylalanine is lower than the two. |
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