| The haploid plants obtained by anther culture can quickly obtain homozygous diploids after treatment with colchicine solution,so as to shorten the years needed for breeding and improve the breeding efficiency.In this experiment,four varieties of Chaotian pepper S48,Fengling pepper,cherry pepper and multicolored pepper were used as experimental materials to study the effect of material genotype on anther culture,according to the relationship between the appearance,morphology and anther color of flower buds and the monocytes of microspores,the material genotype,high temperature(35 ?)and low temperature(4 ?)pretreatment anthers,three kinds of carbon sources(sucrose,glucose,maltose)and concentrations(1%,2%,3%,6%and 9%)and five plant growth regulators(KT,6-BA,2,4-D,ZT,)NAA)to study the effect of embryoid induction rate,so as to provide guidance for anther culture of ornamental pepper.The test results are as follows:1.Observe and measure the external morphological characteristics of flower buds and anthers of four ornamental peppers,and study the relationship between flower buds,anther morphology and microspore development stage by cell observation method.The results showed that the internal mic.rospore development state of the four varieties was inconsistent when the flower bud developed to the same size.However,the color development of anther was consistent,When the front side of anther is 1/3 purple and the back side is 1/4 purple,the microspores were in the mononuclear side stage,and the best inoculation was at this time.2.The anthers of four kinds of ornamental pepper were cultured in the optimized MS medium.The results showed that the embryoid induction rate of Chaotian pepper S48 was as follows:Chaotian pepper S48>multicolored pepper>Fengling pepper>cherry pepper,and the embryoid induction rate of Chaotian pepper S48 was the highest,Up to 24%.3.The anthers of four kinds of ornamental peppers were pretreated at 4?for 1 to 5 days and at 35? for 1 to 9 days respectively,and the effects of two treatments on embryoid induction rate were studied.The results showed that High and low temperature pretreatment had a significant effect on the induction rate of embryoid.The first is that when the induction effect is the same,the low temperature induction time is shorter,and when the embryoid induction rate reaches the highest,the low temperature pretreatment time is generally 2~3 days,and the high temperature pretreatment time is generally 3~5 days.Second,the induction effect of low temperature was better.The induction effect of four kinds of materials(Chaotian pepper S48,wind bell pepper,cherry pepper,multicolored pepper)was 22.65%,2.89%,9.39%and 10.49%higher than that of high temperature treatment,respectively.Therefore,the induction rate of embryoid is closely related to temperature and days,but the effect of low temperature pretreatment is better.The first choice of anther management.4.The orthogonal experiment of different plant growth regulators on embryoid induction of Chaotian pepper S48 showed that the best horizontal combination of embryoid induction was as follows:MS+2,4-D 0.1 mg·L-1+6-BA0.1 mg·L-1+NAA0.5 mg·L-1+ZT1.5 mg·L-1,the induction rate was 46.73%.In orthogonal test,the order of factors affecting embryoid induction rate from large to small was ZT>NAA>KT>2,4-D>6-BA,and only ZT showed significant difference among them.5.Using Chaotian pepper S48 as experimental material,sucrose,glucose and maltose were added to MS medium to observe the number of embryoids under each carbon source concentration.The results showed that within the concentration range of 5 carbon sources(1-9%),when the concentration was 1~3%,with the increase of carbon source concentration,the rate of embryogenesis and emergence increased,and when the concentration exceeded 3%(6%~9%),with the increase of carbon source concentration,the rate of embryogenesis and emergence decreased,and the optimum concentration was 3%.The order of embryoid induction among the three carbon sources was maltose>sucrose>glucose. |
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