人参连作障碍化感作用研究
摘要
连作障碍是制约人参产业化发展的瓶颈。许多研究证实,化感作用是导致人参连作障碍的主要原因之一。鉴于此,本项研究以化感物质组分筛选及化感作用机理为核心,初步探讨人参化感物质对连作障碍的影响,为解决连作障碍问题提供理论参考。主要研究内容如下:
1、栽参土壤提取物活性组分筛选及其化感效应研究
试验采用对栽参土壤水提取液用不同化学试剂萃取和直接采用不同化学试剂提取两种方法,对所得各个组分以人参种子为试材进行生物活性测定,经筛选所得活性组分再以人参幼苗为试材进行化感效应研究。结果表明,栽参土壤提取液不同组分均对人参种子胚根生长有不同程度的影响,化感效应随浓度逐渐增加而增强。采用生物活性较强的栽参土壤水提取液乙酸乙酯萃取相和栽参土壤乙酸乙酯提取液两部分培养人参幼苗,取抑制作用在50%左右的浓度进行培养,来研究活性组分对人参幼苗根系活力和丙二醛含量的影响,测定结果表明,两组活性组分均能促进根系细胞膜质过氧化作用,使膜透性增加,显著降低人参幼苗根系活力。
2、化感物质组分对人参根源愈伤组织生长的影响研究
利用初步确定的人参化感物质-栽参土水提液乙酸乙酯萃取相组分培养人参根源愈伤组织,结果表明,随着培养浓度逐渐增加,对人参根源愈伤组织生长抑制能力越强。通过对愈伤组织生理生化特性研究表明,人参化感物质显著增加愈伤组织中丙二醛含量,降低DNA含量,在培养24h后处理根源愈伤组织DNA含量与对照差异极显著(p<0.01)。通过对愈伤组织解剖结构观察可知,随着培养时间延长,根源愈伤组织细胞壁无序性加强,细胞核逐渐解体消失,使细胞分裂和生长受阻,抑制人参根源愈伤组织生长。
3、化感物质组分对幼苗根系内源激素的影响研究
研究人参化感物质对其幼苗根系内源激素的影响,采用抑制作用在50%左右浓度的人参化感物质培养人参幼苗,分别对培养12h、24h、48h、96h的人参幼苗取样,采用HPLC法进行幼苗根系内源激素(ZT、IAA、GA3、ABA)的测定,结果表明,人参化感物质组分对其幼苗根内源激素平衡有显著影响,在显著降低ZT、IAA、GA3三种促进植物生长的植物内源激素含量的同时显著增高抑制植物生长、促进衰老的ABA含量,使代谢调控系统失衡,从而影响人参幼苗的正常生长发育。
4、人参化感物质对人参幼苗根尖解剖结构的影响研究
人参化感物质具有引起人参幼苗根部细胞膜质过氧化,激素平衡失调和根系活力下降等活性。对于上述生理生化方面的影响必然引起细胞水平上的形态结构变化。利用人参化感物质活性组分培养其幼苗96h后,取其根尖,进行石蜡切片,采用番红—固绿染色,由根尖解剖结构可知,化感物质组分处理后的人参幼苗根尖几个功能部位的细胞发生了明显的大小和排列方面的变化:处理后的幼苗根尖明显变粗,细胞间隙加大,表皮层细胞部分脱落,顶端分生组织和根冠的细胞混乱,这种效应随着处理浓度的增加而增强,且在浓度为40g/ml处理时一部分顶端分生组织细胞的细胞壁形态异常甚至破裂,中柱鞘细胞明显膨胀。
Ginseng continuous cropping obstacle problem has become an important factor which restricts industrialization of ginseng.Many studies show allelopathy is one of the prime reasons which lead to continuous cropping obstacle of ginseng, therefore, the study core was screening of allelochemicals and its mechanism for preliminary study the effects of ginseng allelochemicals on its continuous cropping obstacle,and it could provide highly important theory for resolving continuous cropping obstacle .the main research contents as follows:
1 The study on active constituent screening of extracts from cultivation ginseng soil and its effects of allelopathy
The study applied two methods that used different chemical reagents to extract aquatic extracts from cultivation ginseng soil and directly used different chemical reagents to extract cultivation ginseng soil, and each constituent was done determination of activity with ginseng seeds, and then we researched the effects of higher activity constituents on ginseng seedling.The results showed that extracts from cultivation ginseng soil had effects on radicle growth of ginseng seeds in various degrees,and the allelopathic effects were strengthened with concentration increasing.Both aquatic extracts ethyl acetate phase and ethyl acetate extracts from cultivation ginseng soil manifested stronger inhibition effects,and used them whose concentration were about 50% inhibition effects to culture ginseng seedling .The results indicated both constituents could promote root cell membrane lipid peroxidation and increase membrane permeability, and reduced significantly root activity of ginseng seedling.
2 The study on the effects of allelochemical constituent on ginseng root callus
Ginseng root callus were cultured with allelochemicals.The results showed the inhibition effects were strengthened with concentration increasing.Ginseng allelochemicals could increased remarkably MDA content of ginseng root callus and decreased DNA content,and DNA content of callus between treatment and control groups were significantly different (p<0.01)at the 24th hours after cultivation. We could know from ginseng root callus anatomical structure observation that randomness of cell wall increased and cell nuclear gradually disintegrated and disappeared with prolongation of culture time, and then cell division and growth was inhibited.
3 The study on the effects of allelochemical constituent on ginseng seedling root endogenous hormones
We studied the effects of ginseng allelochemicals on its seedling root endogenous hormones, and used them whose concentration were about 50% inhibition effects to culture ginseng seedling, and sampled at the 12th hours, 24th hours, 48th hours and 96th hours respectively and determined the content of seedling root endogenous hormones (ZT, IAA, GA3 and ABA) by HPLC.The results indicated the effects of ginseng allelochemicals on seedling root endogenous hormones were remarkable,and they significantly reduced the content of endogenous hormones (ZT, IAA and GA3) which were favorable to plant growth, however, they increased the content of ABA which inhibited plant growth and promoted senescence,and then metabolism regulation system was unbalanced and the growth and development of ginseng seedling were influenced.
4 The study on the effects of ginseng allelochemicals on anatomical structure of ginseng seedling root tip
Ginseng allelochemicals could lead to its seedling root cell membrane lipid peroxidation and hormone disequilibrium, and they also could decrease root activity of seedling.They must induce cell morphological structural change due to the effects of allelochemicals on physiological and biochemical aspects of seedling. We used allelochemicals to culture ginseng seedling and sampled at the 96th hours, and made paraffin section with root tip of seedling.The results showed that the treatment seedling cells of several functional regions took place obvious changes in size and arrangement.The seedling root tip became coarse obviously and intercellular space increased, a part of epidermal cells dropped off and the cells of apical meristem and root cap became unordered. The allelopathic effects were strengthened with culture concentration increasing, and some cell walls of apical meristem became abnormal morphology and even fractured in treatment groups with culture concentration of 40g/ml, and the cells of pericycle were also expansive obviously.
1、栽参土壤提取物活性组分筛选及其化感效应研究
试验采用对栽参土壤水提取液用不同化学试剂萃取和直接采用不同化学试剂提取两种方法,对所得各个组分以人参种子为试材进行生物活性测定,经筛选所得活性组分再以人参幼苗为试材进行化感效应研究。结果表明,栽参土壤提取液不同组分均对人参种子胚根生长有不同程度的影响,化感效应随浓度逐渐增加而增强。采用生物活性较强的栽参土壤水提取液乙酸乙酯萃取相和栽参土壤乙酸乙酯提取液两部分培养人参幼苗,取抑制作用在50%左右的浓度进行培养,来研究活性组分对人参幼苗根系活力和丙二醛含量的影响,测定结果表明,两组活性组分均能促进根系细胞膜质过氧化作用,使膜透性增加,显著降低人参幼苗根系活力。
2、化感物质组分对人参根源愈伤组织生长的影响研究
利用初步确定的人参化感物质-栽参土水提液乙酸乙酯萃取相组分培养人参根源愈伤组织,结果表明,随着培养浓度逐渐增加,对人参根源愈伤组织生长抑制能力越强。通过对愈伤组织生理生化特性研究表明,人参化感物质显著增加愈伤组织中丙二醛含量,降低DNA含量,在培养24h后处理根源愈伤组织DNA含量与对照差异极显著(p<0.01)。通过对愈伤组织解剖结构观察可知,随着培养时间延长,根源愈伤组织细胞壁无序性加强,细胞核逐渐解体消失,使细胞分裂和生长受阻,抑制人参根源愈伤组织生长。
3、化感物质组分对幼苗根系内源激素的影响研究
研究人参化感物质对其幼苗根系内源激素的影响,采用抑制作用在50%左右浓度的人参化感物质培养人参幼苗,分别对培养12h、24h、48h、96h的人参幼苗取样,采用HPLC法进行幼苗根系内源激素(ZT、IAA、GA3、ABA)的测定,结果表明,人参化感物质组分对其幼苗根内源激素平衡有显著影响,在显著降低ZT、IAA、GA3三种促进植物生长的植物内源激素含量的同时显著增高抑制植物生长、促进衰老的ABA含量,使代谢调控系统失衡,从而影响人参幼苗的正常生长发育。
4、人参化感物质对人参幼苗根尖解剖结构的影响研究
人参化感物质具有引起人参幼苗根部细胞膜质过氧化,激素平衡失调和根系活力下降等活性。对于上述生理生化方面的影响必然引起细胞水平上的形态结构变化。利用人参化感物质活性组分培养其幼苗96h后,取其根尖,进行石蜡切片,采用番红—固绿染色,由根尖解剖结构可知,化感物质组分处理后的人参幼苗根尖几个功能部位的细胞发生了明显的大小和排列方面的变化:处理后的幼苗根尖明显变粗,细胞间隙加大,表皮层细胞部分脱落,顶端分生组织和根冠的细胞混乱,这种效应随着处理浓度的增加而增强,且在浓度为40g/ml处理时一部分顶端分生组织细胞的细胞壁形态异常甚至破裂,中柱鞘细胞明显膨胀。
Ginseng continuous cropping obstacle problem has become an important factor which restricts industrialization of ginseng.Many studies show allelopathy is one of the prime reasons which lead to continuous cropping obstacle of ginseng, therefore, the study core was screening of allelochemicals and its mechanism for preliminary study the effects of ginseng allelochemicals on its continuous cropping obstacle,and it could provide highly important theory for resolving continuous cropping obstacle .the main research contents as follows:
1 The study on active constituent screening of extracts from cultivation ginseng soil and its effects of allelopathy
The study applied two methods that used different chemical reagents to extract aquatic extracts from cultivation ginseng soil and directly used different chemical reagents to extract cultivation ginseng soil, and each constituent was done determination of activity with ginseng seeds, and then we researched the effects of higher activity constituents on ginseng seedling.The results showed that extracts from cultivation ginseng soil had effects on radicle growth of ginseng seeds in various degrees,and the allelopathic effects were strengthened with concentration increasing.Both aquatic extracts ethyl acetate phase and ethyl acetate extracts from cultivation ginseng soil manifested stronger inhibition effects,and used them whose concentration were about 50% inhibition effects to culture ginseng seedling .The results indicated both constituents could promote root cell membrane lipid peroxidation and increase membrane permeability, and reduced significantly root activity of ginseng seedling.
2 The study on the effects of allelochemical constituent on ginseng root callus
Ginseng root callus were cultured with allelochemicals.The results showed the inhibition effects were strengthened with concentration increasing.Ginseng allelochemicals could increased remarkably MDA content of ginseng root callus and decreased DNA content,and DNA content of callus between treatment and control groups were significantly different (p<0.01)at the 24th hours after cultivation. We could know from ginseng root callus anatomical structure observation that randomness of cell wall increased and cell nuclear gradually disintegrated and disappeared with prolongation of culture time, and then cell division and growth was inhibited.
3 The study on the effects of allelochemical constituent on ginseng seedling root endogenous hormones
We studied the effects of ginseng allelochemicals on its seedling root endogenous hormones, and used them whose concentration were about 50% inhibition effects to culture ginseng seedling, and sampled at the 12th hours, 24th hours, 48th hours and 96th hours respectively and determined the content of seedling root endogenous hormones (ZT, IAA, GA3 and ABA) by HPLC.The results indicated the effects of ginseng allelochemicals on seedling root endogenous hormones were remarkable,and they significantly reduced the content of endogenous hormones (ZT, IAA and GA3) which were favorable to plant growth, however, they increased the content of ABA which inhibited plant growth and promoted senescence,and then metabolism regulation system was unbalanced and the growth and development of ginseng seedling were influenced.
4 The study on the effects of ginseng allelochemicals on anatomical structure of ginseng seedling root tip
Ginseng allelochemicals could lead to its seedling root cell membrane lipid peroxidation and hormone disequilibrium, and they also could decrease root activity of seedling.They must induce cell morphological structural change due to the effects of allelochemicals on physiological and biochemical aspects of seedling. We used allelochemicals to culture ginseng seedling and sampled at the 96th hours, and made paraffin section with root tip of seedling.The results showed that the treatment seedling cells of several functional regions took place obvious changes in size and arrangement.The seedling root tip became coarse obviously and intercellular space increased, a part of epidermal cells dropped off and the cells of apical meristem and root cap became unordered. The allelopathic effects were strengthened with culture concentration increasing, and some cell walls of apical meristem became abnormal morphology and even fractured in treatment groups with culture concentration of 40g/ml, and the cells of pericycle were also expansive obviously.
引文
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