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本文对棉花叶片NR活力的体外分析方法作了两点改进:1)组织匀浆改加提取缓冲液于水浴中,为先在干冰中磨成粉状后再加提取缓冲液溶解;2)匀浆后增加离心沉淀步骤。结果使硝酸还原酶在匀浆中得到充分的保护,完全排除了棉花体内色素对比色的干扰,进一步减少了酚类及其他物质对酶的抑制作用,因此 NR 活力大大提高,约为改进前方法的15倍。用改进的方法对8个陆地棉品种和4个栽培棉种(每种用1个品种作代表),进行 NR 活力的分析、观察,发现陆地棉品种间 NR 活力有明显的差异,同一品种在不同生育期中也有不同,酶活力的这一变化与品种的生长势和对外界氮肥的吸收利用能力有正相关关系。在4个供试栽培棉种中,海岛棉的 NR 活力最高,陆地棉最低,亚洲棉和非洲棉居中。结果还表明4个栽培种的叶绿素含量、光合强度和可溶性糖等生理指标与 NR 活力一致的趋势。
In this paper, we made two improvements to the method of in vitro analysis of NR activity in cotton leaves: 1) adding homogenate to the extraction buffer in a water bath, grinding it into powder in dry ice and then adding extraction buffer to dissolve; 2) Slurry to increase the centrifugal precipitation step. As a result, the nitrate reductase was fully protected in the homogenate, completely eliminating the interference of cotton color contrast color, further reducing the inhibitory effect of phenols and other substances on the enzyme, so the NR activity is greatly improved, which is about the improvement of the former method 15 times. By using the improved method, we analyzed the NR activity of 8 cotton cultivars and 4 cultivars cotton cultivars (each represented by 1 cultivar), and found that there was a significant difference in NR activity between upland cotton cultivars There are also differences in different growth stages. This change in enzyme activity has a positive correlation with the growth potential of cultivars and their ability to absorb and utilize nitrogen fertilizer from the outside world. Among the four cultivated cotton cultivars, the highest activity of NR was found in the island cotton, the lowest in cotton and the highest in Asian cotton and African cotton. The results also showed that the four cultivars chlorophyll content, photosynthetic intensity and soluble sugar and other physiological indicators consistent with the trend of NR activity.