【作者】 张玲；

【导师】 吴国荣；

【作者基本信息】 南京师范大学 ， 植物生理生化， 2008， 硕士

【摘要】 本论文以盐京九号水稻(Oryza sativa L.Yanjing 9)为材料,研究了其抑制铜绿微囊藻(Microcystis aeruginosa)生长的现象及其抑藻的机理,提取分离并鉴定其中的抑藻物质。本论文分三部分:(1)水稻种植水对微囊藻的抑制作用:处理得到种植水(RCW),以其配置BG11培养液,作铜绿微囊藻的培养试验。结果表明RCW抑制藻细胞生长,促进藻细胞黄化死亡。实验过程中藻细胞的生理指标都发生变化:藻细胞生长量和叶绿素含量显著降低;可溶性蛋白含量和SOD酶活性呈先升高后下降趋势。(2)经高温(121℃,20min)处理的和新鲜的秸秆水浸液有相同的抑藻效果,质量比(秸秆/藻液)越大,抑藻效果越明显;呼吸速率和SOD酶活性先升高后急剧下降。秸杆经物理粉碎(0.1mm,0.3mm)后作实验,结果显示相同质量比试验组,粒度越小的试验组抑藻效果越好。曝气处理时间为1d、1w和2w试验组对微囊藻有抑制作用,而处理时间为3w和4w试验组对微囊藻没有明显抑制作用,部分生理指标和固体滤纸片培养也呈现出相应的变化。(3)水稻种植水(RCW)和水稻秸秆水浸液(RSAE)的抑藻物质作初步分离和鉴定:用石油醚、乙醚、乙酸乙酯等有机溶剂依次对RCW和RSAE萃取,各萃取液经浓缩处理,滤纸片法作固体培养基的抑藻实验。结果表明乙醚萃取液的抑藻圈最大。进一步分离乙醚萃取液得到有机酸、酚类物质、中性物质和碱性物质,并作滤纸片试验得知:有机酸组分抑藻效果最明显,酚类物质次之。对RCW和RSAE乙醚萃取物的有机酸组分分别作气-质联用(GC/MS)分析鉴定,RCW组的有机酸组分中有12-甲基十三酸(Tridecanoic acid,12-methyl)、棕榈酸(Hexadecanoic acid)和18烷酸(Octadecanoic acid)等。RSAE组的有机酸的组分中检测出12-甲基十三酸(Tridecanoic acid,12-methyl)、棕榈酸(Hexadecanoic acid)、18烷酸(octadecanoic acid)、苯乙酸(Benzeneacetic acid)和1-甲氧基-13烷酸(Methoxyacetic- tridecyl acid)等。两者的有机酸总离子流图谱很相似,各峰的图谱位置也很接近,表明其具有很高的同源性。图谱中还有很多峰暂时未能鉴别出来,有待进一步的分析。更多还原

【Abstract】 This paper was studied the inhibition of Oryza sativa L.Yanjing 9 on the growth of Microcystis aeruginosa and its inhibitory mechanism.The algae-inhibition compounds were extracted,separated and identified.There were three parts:(1)The study about rice culture water(RCW)inhibited the growth of M. aeruginosa:The RCW was collected and mixed into nutrient solution.Then the RCW was used to cultivate M.aeruginosa.The results indicated that:the growth of M. aeruginosa was obviously inhibited.The cell of M.aeruginosa turned to yellow and died.In the experiment,the physiology index of M.aeruginosa changed:the mass growth and chlorophyll content descended observably.The soluble protein content and SOD activity increased firstly,then decreased as the time went on.(2)The inhibitory effect with the aqueous extract of fresh rice straw was the same as the effect with the aqueous extract of sterilized rice straw.The higher rate of quality(rice straw/water),the more obvious the inhibitory effect was.The respiratory rate and SOD activity decreased at first and then increased rapidly.When the straws were smashed into different particle(0.1mm,0.3mm),the particles were used to cultivate M.aeruginosa.The results indicated that:the smaller particles had better algae-inhibition with the same rate of quality.The straw which treated with oxygen degradation time(1d,1w,2w)could inhibit the growth of M.aeruginosa and the straw of 3w or 4w group had no effect.Some physiology index of M.aeruginosa made the relative phenomena.(3)The RCW and rice straw aqueous extract(RSAE)were separated,purified and identified.The two groups were extracted by petroleum ether,ether and ethyl acetate respectively.Then these extracts were concentrated.The extracts were used to do the experiment of the inhibitory effects on M.aeruginosa(by the method of filtering-paper tests on solid medium).The results showed the filtering paper filling with ether extracts shaped the biggest algae- inhibitory ring.The ether extracts further separated into organic acid,phenols substance,neutral substance and alkaline substance and did the experiment of filtering-paper tests on solid medium.It found that organic acid could shape the biggest algae-inhibitory ring and phenols substance was the next one.The organic acid of RCW and RSAE were made an analysis of GC/MS.The analysis showed that there were Tridecanoic acid-12-methyl, Hexadecanoic acid and Octadecanoic acid in RCW;Tridecanoic acid-12-methyl, Hexadecanoic acid,Octadecanoic acid,Benzeneacetic acid and Methoxyacetic acidtridecyl acid in RSAE.In GC/MS analysis,electron fiow image of two groups(the organic acid of RCW and RSAE)were similar and it explained the two organic acids were comparability.A lot of substants could not be identified and it must be separated in the next step.更多还原