一种蜗牛提取物对花粉破壁方法的研究

Study on cytology of pollen-wall broken by a snail in the extracts

成果背景:花粉里含有丰富的营养成分,被称之"植物微型营养库",但因花粉壁的特殊结构,物理、化学及生物三类七种方法均难以进行破壁,影响了开发利用,即使加温至300℃,也难以使花粉释放出营养成分。方法:本项目受蜗牛于下雨天活动并对植物叶子进行蚕食的启发,捕捉蜗牛经超低温(-45℃)冷冻粉碎,加磷酸盐(p H6.5,0.05 mol)缓冲液提取,离心去沉淀物,取清液,调p H至5.6后,沸水保温至浑浊即流水冷却至常温再离心,放冰箱存放待用;取厨房所收集废蛋壳加1%Na Cl溶液提取,依前述提取蜗牛提取物方法制蛋壳蛋清提取物,放冰箱保存;取沉淀物与前沉淀物作饲料。花粉用1∶1之比例加生理盐水浸提温润后,分别加入前者两个动物提取物破壁,杀菌搅拌,完成破壁灭菌。结果:反应30分钟后显微镜和电镜观察表明,破壁率为97%以上,外表呈明显孔洞或裂沟,紫外分析表明260 nm、280 nm波长吸收值均比破壁前提高30%以上;镜检反应液含菌数<10个(以大肠杆菌数计,每ml数)反应系统混合物进行固液分离,用于食品加工或制作保健品。

[Achievement＇Background]The pollen is rich in nutrition so that it is called the ＂miniature plants＂ nutri- tion library＂, but because of the special structure of pollen wall, seven methods from physical,chemical and biolo-gical are difficult to break the wall thoroughly . It has effects important development and actual utilization. Even if it is heated to 300℃,nutrition is difficult to release from pollen.[Methods] The natural phenomenon that snails eat plants during the rainy days inspired us to raise a new way of dissolving the cell wall. In this project, first, we will reduce snails into particle sizes through cryogenic grinding at a temperature of minus 45℃ and mix it with buffer solution （phosphate solution pH6.5, 0.05mol） to meet the extract requirements. Then, we will get the clear supernatant by through centrifugation, and we should adjust the pH of clear supernatant into 5.6. After that, removing the precipitate we keep the solution in boiling water （100℃） until it gets turbid. Then, separate the liquid by centrifuge again after it restores normal tempera- ture under the flowing water. Finally, We save it into the firdge to use. We can also use the mixture of eggshell and NaCl solution （1%） as materials to get the eggshell extract in the same way. After all these procedures, keep these two solutions refrigerated. Now, we can start the procedures of dissolving the cell wall. First, soak the pollen in the mixture of 1 part pollen solution and 1 part normal saline solution. Then, add snail extract and the eggshell extract in the part pollen solution and 1 part normal saline solution. Then, add snail extract and the eggshell extract in the pollen solution and 1 part normalsaline solution. Then, add snail extract and the eggshell extract in break between these two operations. Mix and sterilize the solution, and finally, we can get the job done.[ Results] the results showed that after 30 min of reaction microscope and electron microscope, the broken rate is above 97%, the appearance of a clear hole or fissure, UV analysis showed that 260nm and 280nm long absorption value than broken before more than 30%; microscopic reaction solution containing bacteria 〈10 （in Escherichia coli, each ml number） reaction the mixture is solid-liquid separation system, used for food processing or production of health care products.