无菌猕猴桃种子采集方法研究

Obtaining aseptic seeds of kiwifruit

无菌的猕猴桃种子是猕猴桃胚乳培养、实生苗微嫁接等技术的基础材料,利用消毒剂灭菌是常用的无菌种子采集手段,应用最广泛的消毒剂为升汞(mercuric chloride)和次氯酸钠(NaClO)。为了避免使用消毒剂,该研究提出了一种新的无菌猕猴桃种子采集方法——无菌搅拌法,同时为探索其准确性和应用性,比较了0.2%升汞灭菌20 min、10%次氯酸钠灭菌20 min、无菌搅拌法三种方式采集无菌猕猴桃种子的效果,并对种子萌发和幼苗形成的影响进行了研究。结果表明:无菌搅拌法、0.2%升汞灭菌20 min是稳定有效的无菌猕猴桃采集方式,10%次氯酸钠灭菌20 min的采集效果不稳定;在相同的时间内,无菌搅拌法的种子发芽率最高,为89.90%,但发芽势较低,均可正常成苗; 10%次氯酸钠灭菌20 min的发芽率次之,与无菌搅拌法的种子无显著差异,发芽势和成苗率最高,分别为47.47%和67.86%,且有打破猕猴桃种子休眠的作用,整体作用类似于赤霉素(GA_3)浸种的效果; 0.2%升汞灭菌20 min对猕猴桃种子的萌发有抑制作用,各项指标均显著低于无菌搅拌法种子,且生长缓慢。此外,无菌搅拌法是物理处理,对种子、操作人员、环境均无害。这证实了无菌搅拌法的实用性和优势,发现了次氯酸钠在打破猕猴桃种子休眠方面的作用,为其它同类型果实的无菌种子采集提供了参考。

Aseptic seeds of kiwifruit is fundamental tested material of endosperm culture, micro-grafting and other researches. It is a usual way to gather aseptic seeds to use disinfectant to sterilize. Among them, mercuric chloride and NaClO are the most widely used disinfectants. This article showed that aseptic stirring method is a new method to obtain aseptic seeds of kiwifruit. In order to explore accuracy and applicability of aseptic stirring method, kiwifruit seeds were used to study the effects of three methods, including 0.2% mercuric chloride sterilization for 20 min, 10% NaClO sterilization for 20 min, aseptic stirring method, and their effects on seed germination and plant growth. The results showed that aseptic stirring method and 0.2% mercuric chloride sterilization for 20 min were an effective and reproductive way for gathering aseptic seeds of kiwifruit. Oppositely, gathering aseptic seeds of kiwifruit which in 10% NaClO sterilization for 20 min was unstable. The results also showed that the three methods were effective, and at the same time, seed germination rate of aseptic stirring method was the highest. And the rate was 89.90%, but its germination energy was lower, sprouts could grow healthily, germination rate of seeds with 10% NaClO sterilization for 20 min was the second place, but there was no significant difference in germination rate of seeds between 10% NaClO sterilization for 20 min and aseptic stirring method, its germination energy and seedling rate were the highest which were 47.47% and 67.86% respectively, and could give the seed a premature start and also could break effectively the dormancy of kiwifruit seeds like GA 3 . And 0.2% mercuric chloride sterilization for 20 min inhibited germination of kiwifruit seeds, the various index was significantly lower than aseptic stirring method, sprouts grew slowly, most of the seeds could not develop into young seedlings within the allotted time. In addition, aseptic stirring method was a physical method, and it was harmless for seeds, workers and environment. The study indicated that the practicality and advantage of aseptic stirring method, and found the ability of NaClO to break the dormancy of kiwifruit seeds, which provided a reference for the obtaining aseptic seeds of other same type of fruits.