Research Of Factors Affacting Embryoid Induction In Pepper Anther Culture

Pepper (Capsicum annuum L.) because of its high nutritional value, and cooking style is varied, therefore, widely grown in our coutry, is a kind of important vegetable crops. Pepper belongs to often cross-pollination crops, natural inspectional is higher, so using traditional breeding methods as normal cultivating pepper, not only time-consuming, laborious and breeding efficiency is low. And the use of anther culture can get pure line in short period of time, thus shortening the breeding cycle, improve the efficiency of breedin, save a lot of manpower and resources. Currently, pepper anther culture has no effective general can directly obtain the embryoid bodies technology system. This study is on the basis of predecessors’research, through a series of factors affecting pepper anther culture embryoids induction and embryoids seedling technology exploration and exploration studies, make pepper anther culture system to further optimize, aims to establish a simple and efficient the system of pepper anther culture. The results of the study are as follows:1. Relations with pepper microspore developmental stages between flower buds external morphological charcteristics Used six different genotype pepper as the test materials, study the relationship between pepper microspore developmental stages with bud external morphological characteristics. The results showed that:there is a certain correspondence between pepper microspore developmental stages and their bud external morphological characteristics and anther color. When the microspore of tested pepper materials in late-uninucleate, petals twilight, calyx and petals isometric or petals slightly longer than calyx, anthers top is light purple, and purple partially accounted for the whole anther 1/4 to 1/3.2. Research of factors affacting anther culture of pepper Used thirteen different genotypes pepper as test materials, research on influencing factors of anther culture of exogenous hormones, carbon sources(sucrose, maltose, glucose), low temperature pretreatment, exogenous organic additives, activated carbon and plant flowering period. Studies showed that:(1) All the tested materials can induce embryoid, there are eight genotypes may obtained robust regenerated plants. Genotype is the key factors of limiting pepper anther culture for inducing embryoid, different test material have different embryo rates, which ’G201’of the embryos was highest rate, reached 31.76%. (2) Different exogenous hormone treatment:NAA is more suitable for anther culture of pepper than IAA, and concentration of NAA and KT in 0.5:1.0 is best. (3) Different carbon source:30g/L of sucrose than 30g/L of maltose is more suitable for anther culture of pepper. (4) Different pretreatments:4℃ cold temperature pretreatment for 1-3d is advantageous to the embryoid induction, handle 2d the embryo rate is highest. (5) Different exogenous additives treatment:beneficial embryoid was 50μmol/L of AgNO3 and activated carbon 4g/L. (6) in early-bloom stage and full-bloom stage buds can be induced into embryoid, the early flowering buds is more suitable for anther culture of pepper.3. Pepper plant regeneration induced rooting and transplanting Used eight genotypes regenerated plants obtained in test as test materials, researching on pepper regenerated plants rooting and acclimatization. The results showed that:The same species of pepper in different hormone concentration treatment, the rooting inducing rate is different, different genotypes in the same hormone concentration treatment the rooting inducing rate also have differences. And the most suitable culture medium for pepper rooting is 1/2MS+0.15mg/LIBA+30g/LSucrose+8g/LAgar, rooting rate up to 100%. Transplanting regenerated plants after rooted survived or not is the last one of the key technologies used in the production of pepper. This article introduces pepper regeneration plant transplanting of domestication, substrate preparation, taking seedlings, disinfection and transplanting technology.4. Ploidy identification of regeneration plant Used the regenerated plants obtained in test as test materials, using the root tip chromosomes identification method to identify its ploidy. The studies showed that:five regenerated plants of the test material’G201’were haploid plants, three were diploid. Three regenerated plants of’G203’had two haploid and one diploid. The regenerated plant of’G205’,’G208’ and ’G213’ was haploid plant. Whereas the regenerated plant of ’G210’ was diploid. The each two regenerated plants of ’G211’ and ’G212’ all was one haploid and one diploid. The identification results showed that the frequency of haploid was 63.16%.