In Vitro Culture And The Analysis Of Genetic Polymorphisms Via ISSR Markers Of Rosa Hybrida

Rose(Rosa hybrida L.) is one of ten famous flowers in China which has a long cultivation history,it has a good name 'queen of flowers'and is popular with people all over the world because of its rich varieties,shining flower color,elegant flower shape and puzzled fragrance,blooming through year.Roses have been widely planted in our country and play an important role in urban greening.This experiment mainly studied the micripropagation,regeneration and application of ISSR markers in genetic polymorphysims of Rosa hybrida.The main results of this study were introduced as following:1.The system of micropropagation of three cut roses and eight miniature roses was established.The results showed:the best micropropagation basal medium was MS, propagation can be carried out on the medium contains lower concentration of NAA (0-0.1mg/L) and BA(0.5-3.0mg/L);stem segment as explant displayed better propagation effect than shoot tip;the highest propagation coefficient was 6.17 of 'Bejing2',the lowest was 2.73 of'Baiminong',the propagation coefficient was only 2.57 of'golden sun'and'Bingjing2'with the stem segment as explants.2.The rooting and transplanting of several cutivars was analyzed.The most suitable basal medium was 1/2MS,the average number of root was14.67 and the average length was 2.53cm in the medium supplied NAA0.1mg/L.After transplanted,its survival rate reached 99%,in vermiculites it grew more stronger than others.But for 'Beiijing6','Golden sun'and 'baxinuo'displayed better significantly on NAA0.1mg/L than 0.05mg/L.3.The regeneration of rose cultivars was optimized.The basal medium was 1/2MS medium supplied with glucose30g/L,Gel2.5g/L,AgNO₃(10mg/L),CH(100mg/L),The best hermone combination of 'Beijing6' was NAA 0.01mg/L and TDZ 1.0mg/L,the regeneration rate is 76%,and on NAA0.01mg/L,TDZ0.5mg/L,NAA0.01mg/L,TDZ2.0mg/L,NAA0.05mg/L,TDZ0.5mg/L,the regeneration rate all higher than 70%;the regeneration rate of 200660 reached 79.2%on NAA0.05mg/L,TDZ1.5mg/L,but low in other medium,we find the regeneration rate of 'Beijing2'and'Beijing5' both displayed best on NAA0.01mg/L,TDZ1.0 mg/L,reached 59.7%and57.3%respectively.Moreover, we tried to compare other 14 cultivars regeneration ability on NAA0.01mg/L,TDZ 0.5mg/L.The highest regeneration can rech 53.3%,then 41.0%,39.0%,6 cultiivars' generation rate is 0.4.The genetic diversity of 62 rose cultivars was analyzed using ISSR marker.From 38 primers,we selected 8 primers,by which 116 clear,reproducible bands was produced, 113 are polymorphic.All individuals can be distinguished with these markers,whieh reveals the level of Polymorphisms of the roses and average 13-16 bands were amplified with one primer.These 116 markers were used to cluster the 62 rose cultivars.The results suggest classification of rose using ISSR data generally supports the traditional classification.