| Pollen encapsulates an abundance of genetic massage for plants and plays a significant role on plant germplasm and crossbreeding. Pollen exposures to environment and is suspectible to many factors. Oil palm pollen has a very short life span in nature which results in difficulties for manual pollination of oil palm.However,under the natural conditions, the life of oil palm pollen is very short, which is not conducive to natural pollination and also brings great inconvenience to manual pollination and crossbreeding.Therefore, research on collection, preservation and utilization of oil palm pollen is of concern to both plant science and agriculture industry. The study conducted two-year field trials and laboratory experiments on pollen collection, preservation, viability test and manual pollination for oil palm tree. Centrally, the study used real-time PCR technique to analyse gene expression during pollen germination.Three types Pisifera, Dura and Tenera were sourced from oil palm germplasm nursery in Wenchang, Hainan of China, a tropical region for oil palm growth. Fresh pollens were hand-harvested for experiements and were tested in different media as well as different temperatures and moistures for germination. According to other reports, sucrose, MS, boric acid and potassium ion stimulated the pollen germination and pollen tube growth under a certain concentration. and the optimum temperature for germination in vitro was seen 31?.And the optimal period for culturing oil palm pollen was observed three hours; For Dura and Pisifera the ingredients of suitable media for pollen germination were comprised of 0.01 g/mL sucrose, 0.05% MS and the absence of boric acid and potassium, subsequently,the rate of the pollen germination was seen up to 93%.Quantitative Real-Time Polymerase Reaction (qRT-PCR) is commonly used for analysing gene expression. The technique features on high-sensitivity, high-resolution,whereby which provides the sequences informative,accurate and reliable. This project applied real-time PCR to analyse gene expression of oil palm (Elaesi guineensis Jacq.)during pollen germination. Three varieties of oil palm were investigated, including Pisifera,Dura and Tenera (You need give general characters of the three varieties here, otherwise,the meaning is incomplete.). The relatively constant 'housekeeping' genes were selected as control,analysing gene expression on quality and quantity pre-and post- germination. Four genes were selected,i.e. OP_Pollen-1,OP_Pollen-2,OP_Pollen-3,OP_Pollen-4. Each gene demonstrated the same trend between the three varieties, and the different genes showed different trends, in particular OP-Pollen-1 and OP-Pollen-4 increased gene products post-germination, OP_Pollen-2 barely changed, and gene products of OP_Pollen-3 was higher pre-germination than post-germination. Study on gene expression of oil palm pollen was developed by real-time PCR as the technique delivers unique and precise results.This study is a Master project which has been conducted for two years. The study investigated three varieties of oil palm widely grown in the tropical growing regions, on the pollens regarding germination conditions, germination mechanism and germination gene expression. Importantly, the study employed the technique real-time PCR to analyse gene expression,select control genes and examine gene functions and thus made a progress for study on pollen germination of oil palm. The significance of the study has drawn many researchers' attention from academic, industry alike and molecular mechanism of oil palm pollen germination. |
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