Study On Sex Differences Between Male And Female Plants Of Brucea Javanica (L.) Merr

Brucea javanica (L.) Merr., is one of the shrub or small tree of Simaroubaceae., which mainly distributed in Fuji an, Taiwan, Guangdong, Guangxi, Hainan, Guizhou and Yunnan provinces. Its mature fruit as an important medicine in southern China can clear heat, detoxicate and defend against dysentery. Brucea javanica emulsion injection, oral emulsion, soft capsule formulation,with Brucea javanica oil as the raw material, were widly used in the clinical treatment of various cancers, leukemia and infectious skin diseases. In addition, in the field of biological protection, Brucea javanica was also a new green pollution-free efficient pesticide.Brucea javanica is a dioecious plant, female plants can bear fruit after flowering. However, it is difficult to distinguish their sex from morphological characteristic before reaching full growth, which has brought some challenges to the large-scale breeding and seedlings planted. Sex identification is able to help to control the ratio between male and female plants, reduce planting costs and improve yields. Sex identification of Brucea javanica in this study includes morphology identification, chemical staining, physiological test and isozymes analysis, the aim is to select the most effective method of sex identification, solve breeding problems and speed up the process of breeding technology. The main contents are as follows:1The review of domestic and foreignAfter reviewing lots of literature, author summarized the general situation of Brucea javanica, including the germplasm resources survey, herbal medicine research, biological characteristics, chemical composition, pharmacology, biological protection and tissue culture. And explained the reasons for the sex differentiation of plants, mainly affected by the factors of genetic material, environment and hormones.2The research of morphological identificationInvestigated leaves length, terminal leaflets length, terminal leaflets width, terminal leaflets index, lateral leaflets length, lateral leaflets width, lateral leaflets index, intersection angles and internodes of female and male Brucea javanica in different areas. By the method of independent samples test, the result showed that there was significant difference among leaves length, terminal leaflets width, lateral leaflets width, terminal leaflets index, lateral leaflets index of female and male Brucea javanica. Principal component analysis was uesd to screen out the main impact indicators, Through principal component analysis, leaf-shaped part as the main impact indicators are screende out. Finally, the classical discriminant function and Bayes discriminant function were established, Bayes discriminant function was more accurate than the classical discriminant function, discriminant functions for mature Brucea javanica were as follows:F1=-67.128+lateral leaflets index×71.673+lateral leaflets width×7.126+terminal leaflets index X96.53+terminal leaflets width×2.063+leaves length×0.696(1)F2=-82.544+lateral leaflets index×70.094+lateral leaflets width×8.371+terminal leaflets index×115.964+terminal leaflets width×2.174+leaves lengthX0.726(2)3The research of chemical staining differencesBromothymol blue and methyl red staining could show color differences between female and, male. Bromothymol blue staining after8h-10h, the female plants were russet, male plants were dark green. Methyl red staining after40h, male plants were scarlet, female plants were yellow-brown. In the dyeing process, the female plants changed color faster than male plants, bromothymol blue staining spent less time than methyl red. In addition, cresol red staining was no significant difference on the color reaction.4The research of physiological test differencesTracking and detecting the changes of content and activity levels of soluble sugar, soluble protein, chlorophyll, MDA, POD and CAT of Brucea javanica within six months, the results showed that the female soluble sugar content was significantly higher than that of male in April, Male soluble protein content were higher than female plants, and revealed the significant difference in September. Male photosynthetic capacity was stronger than female, chlorophyll content in both showed a highly significant and significant differences in June to August. MDA content between male and female reached the highest levels in the summer, but the content of dynamic changes showed no significant difference in half a year. Female POD activity was higher than male in general, both activity had an extremely significant differences in July and August. The changes of CAT activity was stable relatively, there was significant differences between male and female in May.5The research of isozymes analysisPOD isozymes and PPO isozymes of mature plant leaves and buds were significant differences between male and female, POD zymogram of leaves were as similar as buds, male had two bands and female had three bands, POD-1could be the specific band of female plants. PPO isozymes had the different expression form between leaves and buds, male had five bands in leaves and PPO-3as its specific band, female got five bands also and PPO-1as its specific band. In buds, the male plants only showed one band, while female showed two bands and got the PPO-1as its specific band. Identification of unknown sex seedlings could be identified from the number of bands, POD isozyme of male seedlings with two bands and female seedling with three bands, PPO isozymes of male seedlings with one band and female seedling with two bands.