Study Of Aseptic Culture Of Heterodera Glycines And Meloidogyne Incongnita

Four kinds of seeds,soybean,wheat,tomato and cabbage were steriled by NaClO or HgC l₂.Thus steriled vitro roots were obtained.Then H.G lycines,M.incongnita,H.filipjevi were steriled.The steriled cultivation of H.Glycines,M.incongnita and H.filipjevi was achieved by inoculating eggs or the second-stage juveniles.The results were as follows:For soybean seeds,the best steriled method was:The seeds were washed in water for 30 min and sterilized with 5%NaC lO for 20 min under steriled condition.Then soybean seeds were rinsed with sterilized water and peeled.The pollution rate was 11.11%and the germination rate was 83.33%.The 2³ cm long soybean root tip was transferred to B-5 and MS medium under aseptic conditions.Compare with the growth on MS medium,The soybean root tips which on the B-5 medium were grown better.The lateral roots were more stronger and did not appear browning phenomenon on the B-5 medium.For H.Glycines,the best steriled method was method 3:1%NaC lO steriled cysts 12 min.The pollution rate was 5.23%on MS medium.Inoculated by steriled soybean cysts and then 23 petri dishes of soybean roots were cultured at 26?under dark conditions for 34days.Two soybean cysts appeared on two petri dishes.Followed by the method 2:3%NaClO steriled soybean cyst 4 min+1%NaClO steriled eggs for 4 min,and the pollution rate of the eggs on the MS medium was 24%.Inoculated with sterile eggs and then 20 petri dishes of soybean roots were cultured at 26?for 25 days.11 soybean cysts appeared on three petri dishes.Followed by method 4:0.5%NaC lO steriled soybean cysts for 10 min,0.5%NaC lO steriled second-stage juveniles for 4 min,and the pollution rate of second-stage juveniles on MS medium was 17%.Inoculated with steriled second-stage juveniles and then 20 petri dishes of soybean roots were cultured at 26?for 20 days.4soybean cysts appeared on one petri dish.About 10 days the cysts became brown colour.Brown cysts were transfered to new soybean roots.4 so ybean cysts appeared on three petri dishes after 28 days.For wheat seeds,the best steriled method was:1%NaC lO steriled wheat seeds 20 min+3%carbendazim?75%wettable powder?soaking 20 min under steriled conditions,and the pollution rate was 3%,and the germination rate was 40%.Transfer the 2³ cm long wheat root tip?Nongong 211?to MS+0.5 mg/L IBA medium was the best method,and the main roots were longer and the lateral roots were stronger.Followed by the transfer of wheat root tip?Nongong 211?to MS medium.Followed by the transfer of wheat root tip?dwarf resistant 58?to MS medium.For Meloidogyne incongnita,the best steriled method was:Tomato seeds and cabbage seeds were washed for 30 minutes.Sterilized with 5%NaC lO for 20 min under steriled conditions,the contaimination rate of tomato seeds was 0%and the germination rate was96%;The contaimination rate of spinach seeds was 0%and the germination rate was 94%.The growth of spinach roots on MS medium was better than tomato roots.The three eggs of Meloidogyne incongnita were steriled with 1%NaC lO for 4 min,and rinsed with steriled water for six times.The eggs were inoculated around the sp inach roots and cultured at 26?under dark conditions.After 11 days,the root knots appeared on the spinach root tips.After50-60 days,a large number of second-stage juveniles appeared on the medium.The contaimination rate was 23%.