| Lycium barbarum L. belongs to the Solanaceae,Lycium,and perennial deciduous shrub. In this research, Lycium barbarum L,as the research object to explore the relationships between the growth of wolfberry fruit(Diameter, Length, Hundred-grain weigh) as well as its polysaccharide content and the main meteorological factors, At the same time, the LBP (Lycium barbarum polysaccharides) were extracted and purified and the variety of antibodies againsted LBP were screened. Using the methods of immunohistochemistry(immunofluorescence and immunogold) to the localization of LBP in different development periods of Lycium barbarum L.fruit. The results can reveal the relationship between the Structure development of wolfberry fruit and the accumulation of polysaccharide content,simultaneously,providing a scientific basis for practical issues such as improving the yield and its active ingredients content1. The fruit growth curve of Lycium barbarum L. in 3 major producing areas belongs to the double"S"curve. Wolfberry fruits from Yinchuan area had their first growth spurt 0-8 days after blooming, and from Baiyin it was 0-12 days after blooming, from Delingha it was 0-16 days after blooming. The slow periods for growth were respectively 8-24 days,12-28 days and 16-32 days after blooming. The second growth spurt took place respectively at 24-34 days,28-40 days and 32-52 days after blooming. The entire growth of wolfberry fruit needed 34 days,40 days and 52 days separately. Average temperature(AT) and average temperature difference between day and night(ATD) were two main meteorological factors that affecting the growth and development of wolfberry fruit. During the whole developmental process, growth rate of fruit from Yinchuan area increased firstly with the rising temperature and reached a peak at 23.71℃,23.93℃ and 23.55℃,then slowed down in spite of the rising temperature. Growth rate of fruit from Baiyin area continually increased with the rising temperature and at 22.99℃,22.16℃and 21.35℃ it slowly reached to its peak. So did the growth rate of fruit from Delingha area whose peak temperature lay whereas in 19.55℃,21.01℃and 20.64℃2. Polysaccharide content in fruits from three areas always showed a trend of Yinchuan>Baiyin>Delingha during developmental process and increased with the fruits’physical improvement. The polysaccharide content increased in slowly phase (Yinchuan 0-24 days, Baiyin 0-28 days,Delingha 0-32days)and rapid phase(Yinchuan 24-34 days,Baiyin 28-40 days,Delingha 32-52 days) Whether it was the fruit growth or the polysaccharide content of the wolfberry fruit from three producing areas, they all increased with the expansion of AT, which turned out to the main affecting meteorological factor to polysaccharide content of wolfberry fruit.3. ELISA screening against polysaccharides of Lycium Barbarum L.was used to select effective 10 primary antibodies (JIM94,JIM8,JIM7,JIM12,JIM13,MAC204,JIM11,JIM19,JIM20,JIM5) from 30 previously generated mAbs. Western blot analysis indicated the presence of four epitopes recognized by the anti-arabinogalactan protein antibodies JIM94,JIM8,JIM13,MAC204 and anti-extension antibodies JIM11,JIM20 and anti-pection antibodiesJJIM5 in fruits of Lycium Barbarum L.The ability of these seven antibodies against of LBP are superior for AGPs.Immunofluorescence localization of LBP showed that LBP accumulate mainly at the exocarp, some parenchyma cells and vascular bundles in the mesocarp.The pericarp cell structure of Ningxia wolfberry were complete and with the highest fluorescence in the first rapid growth phase, fluorescence was reduced with the development of fruit. Gold labelling was mainly present within the region between cell wall of parenchyma and plasma membrane,vacuole membrane and vesicles within the vacuole,chloroplasts,chromoplasts and other plastids within cytoplasm and the membrane of them.We observed that the cell wall structure is relatively complete during the early stage,with the development of the fruit,the cell wall of the parenchyma became loose, plasmolysis appearanced.We observed that the plasma membrane invaginated and vesicles formated. During the second rapid growth phase,the wall of parenchyma cell was disintegrated and disappeared gradually, plasmolysis turned seriously;a large number of invagination was apperared and a large amounts of vesicles informed innside the vacuole. We considered that LBP were produced by the disintegration of the cell wall and transported from the cell wall to other parts of the cell through the form of vesicular transportation. |
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