Factors Affecting Embroidgenesis In Peppee (Capsicum Annum L.) By Anther Culture

In this paper, the convenient and reliable methods of selection of anther culture was determined, and some key factors were studied which affect the anther culture of pepper, such as genotype, low-temperature pretreatment, high temperature induction, the concentration of activated carbon and silver nitrate and hormone in the basic medium, and so on, the major results were as follows:1.There are certain corresponding relationship between the morphological characteristics of flower buds and development stage of microspore in pepper. When the calyx is encased by corolla and the corolla is green and the calyx is longer than corolla and the anther is greenish white, most of the microspore are at the early nucleate stage. When the corolla emerges a little and the corolla is greenish white and the corolla is equal to or slightly longer than the calyx and the anthers have a violet tinge at their ends, most of the microspore are at the late nucleate stage, while the nucleus is situated on one side of the pollen. When the corolla shows up remarkably and the corolla is greenish white or white and the corolla is longer than calyx and the anther are all purple, most of the microspore are at the binucleate stage.2.The effect of genotype of donor plants on anther culture of pepper was significant. The induction rate of anther embryoid is different with different genotype of donor plants, Among the nineteen genotypes for test, seventeen embryo are induced, the rate of success of induction is 89%.To different pretreatment time and additives in medium and different hormone concentration, there are large difference among varieties. The donor plants of highest induction rate is PN3-13, the percentage of embryoid formation is 5.125. P142 only induced two embryoid., under the same conditions,PN3-8-9 and PN1-5 don't give birth to embryoid.3.Low-temperature pretreatment on buds could increase the rate of induction。P120,PN3-13②,P130 give rise to the highest percentage of the rate of inducing the embryoid after 3d low-temperature pretreatment. P140,PN1-4 give rise to the highest percentage of the rate of inducing the embryoid after 5d low-temperature pretreatment, the induction rate of P140 might reach 7.272%.The induction rate of the material which are pretreated by low-temperature in 1～5d increase first and decrease ,the induction rate is the highest, after pretreatment 3d or 5d. The induction rate decreased rapidly, The CK induction rate is very low, even no embryoid. 4.The inoculated anther was treated with 33士1℃high temperature for one day, three days, five days,seven days,nine days and eleven days, respectively. The result shows: after the high temperature treatment, the percentage of the embryoid formation enhances remarkably. After 3d high temperature, with the time of high temperature treatment protracted, the induction rate is also enhancing. The induction rate is the highest after high temperature treatment for seven or nine days, over nine days, the induction rate descended. After treated eleven days ,the rate descended suddenly.5.When three genotype are cultured in the medium whose concentration of AgNO3 is 10,50,100μmol/L,the induction rate of embryoid are enhanced in some degree.In certain extent, the induction rate of embryoid is improved with the increasing of concentration of AgNO3 .When the concentration of AgNO3is 50μmol/L, the induction rate is the highest.After that,the induction rate is decreased with the increasing of concentration.6.When P140,P120,PN3-13,PN1-4 are cultured in the medium whose concentration of activated carbon is 0,0.3 %,0.5 %,1% , PN3-13,PN1-4don't give birth to embryoid in CK medium, but in concentration of 0.3%-1.0%,the embryoid is inducted. The induction rate of P140,P120 is very low, while in concentration of 0.3%-1.0%,it is increased. In concentration of 1 % , the induction rate is the highest.7.Adding suitable hormones in MS medium can increase induction rate of embryoid .In low concentration of 2,4-D medium,the induction rate of embryoid of inoculated four genotype are all improved. after, P120,P132,P134 reach very significant level comparing with the CK. When the concentration of 2,4-D is 0.5mg/L, the induction rate is decreased.8.Adding KT(0.1mg/L) of low concentration to medium can increase the induction rate of embryoid obviously.With the concentration of KT increased , the induction rate of embryoid decrease in different degrees.9.The tests indicate that the induction rate of P140 and PN1-13 improved with the increasing of the supplementation of NAA, when the concentration of NAA reach 1.0ml/L , the induction rate is the highest ,after that,it is reduced rapidly.When the concentration of NAA reach 1.5mg/L, no embryoid can be inducted .It has positive function to anther culture of pepper adding proper NAA.