Induction And Preservation Of Somatic Embryo In Wine Grapevine

The　ovules　of　nine　wine　grapevine　cultivars,　one　table　grapevine　cultivar　and　four　rootstocks　cultivars　were　used　in　the　experiment.　There　were　significant　effect　of　growth　regulator　combinations,basal　media,genotypes　and　embryo　age　on　induction　of　grape　callus.　It　was　studied　culture　condition,　carbon　sources　and　additives　affecting　formation　of　grape　embryoid.The　basal　media　had　effected　on　embryoid　germination　and　plantlet　regeneration.In　addition,the　optimal　medium　and　conditon　which　influenced　preservation　of　EC　and　embryoid　desiccation　were　also　studied.Some　conclusions　were　summarized　as　followed:　1　　The　effects　of　some　factors　on　EC　induction　of　grape　were　investigated　by　using　the　orthogonal　test.　The　results　showed　that　the　best　medium　for　EC　induction　　was　NN+0.5mg/L　2,4-D+1.5mg/L　6-BA(NN3).The　result　of　sifting　basal　media　showed　that　NN　was　superior　to　MS　and　B5.Genotypes　had　strong　influence　on　callus　induction　.The　ovules　of　eleventh　cultivars　had　formatted　callus　and　there　was　a　significant　difference　in　texture　and　quantity　of　callus,only　six　cultivars　formatted　EC　and　EC　forming　rate　varied　considerably　in　different　genotypes,EC　of　Sinsaut　was　induced　in　the　highest　rate　and　EC　of　Heijiamei　was　induced　in　the　lowest　rate.The　explants　of　different　embryo　ages　showed　that　the　rate　of　callus　induction　was　higher　than　75%,which　was　after　anthesis　from　the　eighth　to　the　tenth　week.　2　　The　proliferation　of　EC　was　studied.　The　results　indicated　that　the　average　　growth　yield　and　average　proliferation　multiple　were　as　followed:　Liquid　medium﹥solid　medium﹥shallow　layer　medium.The　average　growth　yield　and　proliferation　multiple　of　liquid　medium　were　up　to　0.3236g　and　7.01　respectively,so　liquid　medium　NN3　was　used　in　the　proliferation　of　EC.　　3　　The　EC　were　transferred　to　NN+0.5mg/L　6-BA+0.03mg/L　NAA(NN2)　and　the　ratio　of　dark　time　to　light　time　was　2　to　1,　which　was　the　better　condition　for　embryoid　　happening.In　the　experiment　of　embyoid　induction,　some　other　conditions　were　investigated.　The　results　showed　that　the　rate　of　embyoid　induction　was　higher　than　50%,　when　NN2　added　with　30g/L　sucrose　and　0.3g/L　CH　or　LH.The　concentration　of　AC　was　from　1.5g/L　to　3g/L,　which　was　beneficial　of　somatic　embyogenesis.　　4　　GS+0.2mg/L　IAA+0.015g/L　penicillin　was　optimum　for　embryoid　regeneration.　The　rate　of　somatic　embryo　germination　was　52.5%,plantlet　regenration　rate　was　33.4%　and　rooting　rate　was　69.8%.　5　　After　preservation　in　NN3　solid　medium　for　about　four　months,　EC　should　be　transferred　to　NN2　solid　medium　and　cultured　20　days　and　then　good　EC　should　be　choosed　to　preserve　in　NN3　solid　medium.　NN3　solid　medium　alternated　with　NN2　solid　medium,which　was　favorable　to　EC　proliferation　and　long　period　preservation.　The　suitable　preservation　condition　for　EC　was　weak　light　or　dark　and　a　temperature　of　4±1℃.Under　this　condition,　the　growth　yield　of　EC　was　relatively　reduced　and　subculture　period　was　prolonged,　so　it　was　beneficial　for　the　preservation　of　EC.　6　　The　rate　of　somatic　embryo　germination　related　directly　to　the　quantity　of　loss　water.　When　the　quantity　of　loss　water　ranged　from　0　to　50%,　with　the　increase　of　it,　the　rate　of　somatic　embryo　germination　could　be　improved.　Especially　,when　the　quantity　of　loss　water　ranged　between　40%　and　50%　,　the　rate　of　germination　could　be　9.4%　higher　than　the　germination　rate　of　those　embyoids　without　desiccation.　Somatic　embryos　were　desiccated　at　RH　30-90%　after　6-192　hours　of　storage,　the　results　showed　that　the　quantity　of　loss　water　of　somatic　embryo　was　slow,the　time　for　desiccation　was　prolonged　and　the　rate　of　surviving　and　germination　was　improved　at　the　higher　RH.　Somatic　embryos　were　desiccated　15　days　at　RH　70%,　which　was　beneficial　of　somatic　embryo　germination.　The　rate　of　germination　was　raised　from　33.5%　non-desiccation　treatment　to　56.2%　with　15　days　desiccation　treatment.