Establishment Of High-frequency Regeneration System From Common Wheat Variety Shannong33of Tissue Culture

For establishing a stable and efficient in vitro tissue culture system, this paper onShannong33and two controls(Kenong199and H269) culturing immature embryos,Comparison of Shannong33and three control materials(Kenong199、Bobwhite and Lunxuan987) tissue culture ability of mature embryo in four kinds of medium, to explore relatedimpact factors of high frequency regeneration system established, and comparison twentywheat varieties to the ability of mature embryo tissue culture, the study obtained the mainfollowing results:1.Wheat varieties (lines) Shannong33, Kenong199and H269immature embryo tissueculture, the highest average inducted rate of Kenong199, up to99.43%; Shannong33was98.11%; the lowest is H261at96.24%; the browning of H261is the highest rate to7.81%,while the browning rate of Shannong33and Kenong199is similar, Kenong199(2.25%),Shannong33(2.48%);The quality callus of the inducting rates: Kenong199(97.20%),Shannong33(95.68%), H261(88.73%). Regeneration and differentiation of the three loworder: Shannong33(84.68%)> Kenong199(72.62%)> H261(64.72%).The number of callusgreen spots per embryos: Kenong199(11.33) highest, the following Shaannong33(10.00),the lowest is H261(5.67).2. When AgNO3of the best concentration was4.0mg/L, for inducting anddifferentiating callus of the immature embryos of Shannong33, inducting rate up to99.41%,renewable differentiation rate to82.35%, the browning rate was1.68%, and under othergroups of AgNO3concentrations total callus is absence of browning; when CuSO4concentration2.5mg/L, the highest rate of differentiation, was84.17%, and the rate ofadding CuSO4to the differentiation medium was the higher than the control.3. Four kinds of wheat varieties Shannong33, Kenong199, Bobwhite and Lunxuan987is selected for the material, under four kinds of induction media high quality callus onaverage inducting rate of relationship：MSSR(97.92%)、MSB5(96.00%)、W14(95.97%)、N6(95.00%); and most produced callus on MSSR and MSB5medium is significantly betterthan on W14and N6medium. Under the differentiation medium on four kinds of callus tissue average differentiation rate were: MSSR(58.30%)>MSB5(45.83%)>W14(42.92%)>N6(36.61%).On the four kinds of medium callus average inducting rate of four materials were:Shannong33(96.50%)>Bobwhite(95.92%)~Lunxuan987(95.92%)>Kenong199(95.67%)，theaverage differentiation rate fromfour kinds of wheat varieties were: Bobwhite(53.78%)>Kenong199(46.08%)>Lunxuan987(42.40%)>Shannong33(41.80%).4. The20kinds of wheat varieties (lines) of mature embryos were cultured on twodifferent types media of MSB5and MSSR., the average inducting rate of the other18kindsof wheat materials have reached more than95%, Where K35(60.50%) andJingdong8(57.46%) of average inducting rate is the lowest, and which twelve of all materialsthe quality of callus performance is much better.Regenerating rate on two kinds of differentiation medium, Bobwhite are the highest,and K35are the lowest; on the differentiation medium of MSB5, only Bobwhite andKenong199of differentiating rate reached up to50%above, respectively55.24%,51.68%,and the rest of regenerating rate is between28.30%and44.59%; in differentiation mediumof MSSR, Bobwhite and Kenong199of differentiating rate were67.18%,60.34%, the other18genotypes is between34.60%and55.82%;Under the differentiation and regeneration medium of MSSR, there are five genotypesof more than50%respectively: Bobwhite(67.18%)、 Kenong199(60.34%)、Shannong138(55.82%)、Lunxuan987(54.39%)and Shannong33(50.17%) are five genotypesacceptor material among20kinds of materials, could make a relatively higher differentiationand regeneration ability.