安徽不同生态区域油茶叶片结构及生理生化特性的研究
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摘要
油茶(Camellia oleifera Abel.)是集经济、生态和社会效益于一体的木本油料树种，近年来被广泛栽植。安徽为暖温带向亚热带过渡的气候类型区，既是油茶适宜分布区之一，也是油茶分布的北缘地带，故影响油茶发展的因素较多。为了更好地促进安徽油茶产业的健康发展，给安徽江淮地区发展油茶提供更多的理论依据，本文选择了皖东、皖西以及皖南三个生态栽培区的21个油茶品种为试材，较系统地研究了安徽不同生态区域油茶叶片解剖结构、酶活性、电导率、苯醇抽提物、纤维素含量等生理生化特性及活性物质的差异，研究结果表明：
1.不同生态类型区的6组油茶4个物种气孔密度的测定结果表明，江西小果、普通油茶（黄山、大别山、凤阳油茶）、博白大果与攸县油茶的气孔密度差异较大且显著，分别为6.50个/mm~2、4.35个/mm~2（普通油茶气孔密度均值）、3.20个/mm~2、2.8个/mm~2，而黄山、大别山、凤阳油茶等普通油茶同一物种间的气孔密度无显著差异。
2.通过对不同生态区域油茶品种叶片解剖结构进行观察测定发现，不同地区间油茶叶片的上下表皮、角质层、栅栏组织、海绵组织厚度等叶片表皮结构差异十分明显，且上述组织厚度由大到小排序为：凤阳油茶>大别山油茶>黄山油茶。由此可证明，凤阳地区油茶品种为适应北方低温、干旱等气候条件，叶片组织在解剖上形成了抵抗逆境环境的结构特征。
3．不同生态区域年平均降雨量与油茶叶片气孔开度和叶片细胞结构的通径分析结果表明：年平均降雨量与油茶叶片气孔长度、气孔个数、气孔开度呈正相关；但与叶片角质层的厚度却呈负相关，且有明显的直接作用。另外，在对不同生态区域油茶叶片气孔开度与叶片细胞结构中的其他各项指标多元逐步回归分析中发现，叶片气孔开度与气孔尺寸有直接正相关关系，但与叶片上、下表皮的厚度关系均为负相关。
4．本文通过DPS数据处理软件对来自三个生态区域的6个油茶物种的叶片细胞气孔开口大小进行聚类分析，结果表明：在遗传距离为1.06cm处，6个油茶种质资源共分为5个类群。类群1包括博白大果（2号）和江西小果（5号）2个物种，它们的样品相似性最大，亲缘关系最为接近。凤阳油茶亲缘关系较远，可能是该油茶长期生长在相对温度较低、降雨量较少的环境下，导致在形态上产生了较大的变异。4号攸县油茶与其他的油茶物种亲缘关系最远，其主要原因可能是物种的不同。
5．通过对不同生态区域油茶植株含水量及电导率测定发现，油茶叶片电导率以皖东地区偏低，但三个生态类型区的差异并不显著。植株新梢、叶片含水量的测定结果表明：皖南>皖西>皖东，即植株含水量与降雨量表现出明显的正相关关系；在同一生态类型区，油茶新梢含水量高于叶片含水量。
6．通过对不同生态区域油茶叶片丙二醛（MDA）及脯氨酸（Pro）含量的测定发现，三地区油茶叶片MDA含量排列顺序为：皖南地区3.227μg/g>皖西地区2.836μg/g>皖东地区2.675μg/g；当年生叶片Pro含量皖南油茶最高，达到1192.963μg/g，依次为皖西和皖东，分别为1158.938μg/g和997.129μg/g，其排序与丙二醛含量相一致。该结果表明：地处油茶分布最北缘地带的皖东地区油茶抗逆性较强。
7．通过对三个生态区油茶SOD、CAT和POD三种酶活性的测定试验结果表明：皖东地区油茶植株的SOD活性高于皖西和皖南；油茶植株CAT活性也是皖东地区高于皖南和皖西地区，但它们之间的差异不如SOD那样显著；与以上相反，油茶植株的POD活性皖东地区为最低（仅为1.5061U/g.min），而皖西和皖南地区均较高，都达到了5.2U/g.min以上。
8．通过对同一生态区域油茶品种野外调查及对其SOD、POD和CAT三种酶活性的进行了比较分析，SOD、CAT酶活性在同一地区良种的含量高低排序中出现高度的一致，分别为：皖东地区凤阳4号>凤阳3号>凤阳2号>凤阳1号>凤阳5号；皖西地区大别山3号>大别山1号>大别山4号>大别山2号；皖南地区黄山1号>皖徽5号>皖宁5号>皖宁2号>黄山2号>皖徽3号>黄山4号。这说明了SOD、CAT两种酶在油茶保护酶系统中的共同作用和核心地位。
另外多重比较分析结果表明，同一地区不同油茶良种之间POD活性差异显著，皖东凤阳2号POD活性最低，只有0.736U/g.min，凤阳1号和凤阳3号POD活性较高且差别不大；皖西仅大别山2号POD活性较低，其它良种POD活性较高，且均在6.762U/g.min以上；皖南油茶良种POD活性为黄山2号和皖宁5号含量较低，但和其他良种POD活性差异显著。
9．不同地区油茶品种的内含物研究结果表明：在所测的油茶试样中，含水量、苯醇抽提物、综纤维素、纤维素和木质素的含量分别为：44.3%-50.1%、10.8%-26.2%、0.28%-0.57%、0.12%-0.25%和0.13%-0.28%。在同一生态类型区，油茶体内仅苯-醇抽提物含量为叶片高于枝条，其余物质含量均是枝条高于叶片。在三类生态类型区中，最北缘（凤阳）油茶林的枝条、叶片苯醇抽提物含量、纤维素及木质素含量最高；含水量（枝和叶）和综纤维素（枝）为最低；其中油茶枝条的综纤维素含量差异极显著(P=0.0084<0.01）、纤维素含量差异显著(P=0.0214<0.05)；叶片综纤维素含量差异显著(P=0.0467<0.05)；由此可见，油茶体内较低的水分，较高的苯醇抽提物、纤维素和木质素含量有利于植株抗寒和抗旱能力的提高。
10．通过用高分辨质谱联用法（HPLC-DAD-MS）对皖西的大别山3号、皖南的黄山1号、皖东的凤阳4号叶片的甲醇和乙酸乙酯混合提取物进行了比较分析，发现生长在三个不同生态区域油茶品种中，大别山3号特有的化合物阴离子有C_(18)H_(24)O_(11)、C_(21)H_(20)O_(11)，阳离子有C_(23)H_(18)N_2O_4、C_(24)H_(30)O_6和C_(12)H_(20)N_4O_5；黄山1号特有的化合物有C_(27)H_(36)O_(12)；凤阳4号特有的化合物阴离子有C_(32)H_(42)O_3，阳离子有C_(16)H_(35)NO_2、C_(18_H_(39)NO_3和C_(20)H_(43)NO_4。三种生态类型的油茶叶片的活性化学物质差别较大，每个地区的叶片内都具有不同的抗性物质。其中皖东地区的凤阳4号阳离子化合物除含有脂肪酰胺类物质，还具有C_(16)H_(35)NO_2和C_(18)H_(39)NO_3等鞘氨醇类物质，鞘鞍醇类物质是一组具有显著抗性的活性物质，这也可能显著提高了处在北缘地区的凤阳4号油茶的抗性。
Camellia oleifera Abel. is woody oil tree having economic, ecological and socialbenefits, which is widely planted in recent years. Anhui province is a climate transitionregion from warm temperate zone to subtropical, which is one of C. oleifera appropriatedistribution area and is on the northern margin of C. oleifera. Thus, the factors affecting thedevelopment of C. oleifera are complicated. To promote the health development of AnhuiC. oleifera industry and to provide more theoretical basis for jianghuai region,21C.oleifera varieties from eastern, western and southern Anhui were selected as experimentalmaterials, and leaves physiological and biochemical characteristics such as Anatomystructures, enzymes activity, conductivity, benzene-ethanol extracts and cellulose content,and the differences of active substances in different ecotypes were studied, the resultsshowed that as follows:
1. Stomatal density of4species,6groups C. oleifera in different ecotypes weredetermined and the results indicated that there was significant difference between C.meiocarpa, C. oleifera (Huangshan, and dabieshan and fengyang C. oleifera), C.gigantocarpa and C.yuhsienensis, with the value of6.50a/mm~2,4.35a/mm~2(mean valueof C. oleifera stomatal density),3.20a/mm~2,2.8a/mm~2respectively. However, there wasno significant difference in stomatal density level between the same species of C. oleifera.
2. Leaf anatomical structures of C. oleifera varieties in different ecotypes weredetermined and the results showed that there was significantly different in leaves upper andlower epidermis, cutin layer, stockade and spongy tissue thickness levels. Moreover, theordering of the above tissue thickness of C. oleifera in different ecotypes was as follows:Fengyang> Dabieshan> Huangshan. Therefore, the results indicated that leaf tissuestructure of Fengyang C. oleifera formed the characteristic structure, and to adapt to thecold, drought and other adverse northern climate conditions.
3. The path analysis between and stomatal opening and leaf cell structure of C.oleifera were carried out and indicated that average annual rainfall in different ecotypeswere positively correlated with leaves stomatal length, stomatal number per unit andstomatal opening, while negatively correlated with leaf cutin layer thickness. In addition, the multiple stepwise regressions between the leaf stomatal opening and the otherindicators in leaf cell structure of C. oleifera in different ecotypes showed that leafstomatal opening was positively correlated with stomatal length, and negatively correlatedwith leaves upper and lower epidermis thickness.
4. leaf cell stomatal opening of6C. oleifera cultivars in three ecotypes were clusteranalyzed by DPS data processing software, and the results showed that6C. oleiferacultivars was divided into5groups, when the genetic distance was1.06cm. C.gigantocarpa (No.2) and C. meiocarpa (No.5) were grouped in Class1, which had amaximum similarity and a relatively close relationship. Fengyang C. oleifera was moredistant from other cultivars, because the cultivar grew long-term at a relatively lowtemperature and low rainfall, leading to a large variation of morphology. No.4C.yuhsienensis was the most distant from the other cultivars; the main reason was the cultivarfrom different species.
5. Water content and conductivity of C. oleifera leaves in different ecotypes wasdetermined and found that leaf conductivity of eastern Anhui was the lowest, while theconductivity in three ecotypes was not different; the ordering of water content wassouthern Anhui> western Anhui> eastern Anhui. Moreover, leaf water content waspositively correlated with rainfall, and water content of shoot was higher than that in leaf inthe same ecotype.
6. leaves MDA and Pro content of C. oleifera in different ecotypes weredetermined and the results showed that the ordering of MDA content was southern Anhui(3.227μg/g)> western Anhui (2.836μg/g)> eastern Anhui (2.675μg/g); leaves Procontent in southern Anhui was the highest, reaching1192.963μg/g, followed by westernAnhui and eastern Anhui, with the value of1158.938μg/g and997.129μg/g respectively,MDA content had the same tendency with Pro content in different ecotypes. The aboveresults indicated that eastern Anhui, which is located in the northern fringe of Anhui, had astrong resisting property.
7. SOD, CAT and POD activities in the three ecotypes were determined andanalyzed that SOD and CAT activities of eastern Anhui were higher than that of southern and western Anhui; there was significantly different in SOD activity level; on the contrary,POD activity in eastern Anhui was the lowest with the value of1.5061U/g.min, whereaswestern and southern Anhui was higher, reaching above5.2U/g.min.
8. SOD, POD and CAT activity in the same ecological region were compared andanalyzed. SOD, CAT enzyme activity in the same area had the same tendency: easternAnhui, Fengyang No.4> Fengyang No.3> Fengyang No.2> Fengyang No.1> FengyangNo.5; western Anhui, Dabieshan No.3> Dabieshan No.1> Dabieshan No.4> DabieshanNo.2; southern Anhui, Huangshan No.1> Wanhui No.5> Wanning No.5> Wanning No.2>Huangshan No.2> Wanhui No.3> Huangshan No.4. This explained the two enzymes ofSOD, CAT playing an important role in the protective enzyme system of C. oleifera.
In addition, multiple comparison analysis showed that POD activity was significantdifference between different varieties of C. oleifera in the same area. POD activity ofFengyang No.2in eastern Anhui was the lowest, only0.736U/g.min, and Fengyang No.1and Fengyang No.3was higher and the difference of POD activity in the two varieties waslittle; POD activity of Dabieshan No.2in western Anhui only was the lowest, POD activityin other cultivars was higher, reaching above6.762U/g.min; POD activity of HuangshanNo.2and No.5in southern Anhui was lower, there was significant difference between theabove cultivars and other cultivars.
9. Leaf inclusions of C. oleifera cultivars from the three regions were studied thatthe range of the content of water, benzene-ethanol extracts, holocellulose, cellulose content,lignin content of all the tested were44.3%-50.1%,10.8%-26.2%,0.28%-0.57%,0.12%-0.25%and0.13%-0.28%respectively. The same material content in different tissueof C. oleifera of the same ecological type was different, including the content ofbenzene-ethanol extracts of leaves was higher than that of branches, and the other materialcontents were quite the contrary.(3) The material content of branches and leaves weredifferent with ecotypes, the content of benzene-ethanol extracts, the cellulose content andlignin content of eastern Anhui were the highest, followed by western and southern Anhui;the water content of eastern Anhui was the lowest; there were significant differences of thecontent of holo-cellulose(P=0.0084<0.01), cellulose content (P=0.0214<0.05)of branches at the level of0.05, with the range of0.84%-2.14%; and there was significant difference inthe content of holo-cellulose(P=0.0467<0.05) of leaves at0.05; there was no difference inwater content and lignin content. In the future production of C. oleifera of northern edge,many different cultivation measures such as improving the content of benzene-ethanolextracts and the cellulose and lignin, reducing water content and holo-cellulose contentwere taken to improve the stress resistance of C. oleifera.
10. Methanol and mixed solvent of choroform and ethyl acetate extracts from leavesof Dabieshan No.3in western Anhui, Huangshan No.1in southern Anhui and FengyangNo.4in eastern Anhui were analyzed with HPLC-DAD-MS. In three ecotypes, DabieshanNo.3had unique negative ions of C18H24O11, C_(21)H_(20)O_(11), positive ions of C_(23)H_(18)N_2O_4,C_(24)H_(30)O_6and C_(12)H_(20)N_4O_5, Huangshan No.1has unique compounds of C_(27)H_(36)O_(12);Fengyang No.4had unique negative compounds of C_(32)H_(42)O_3, positive ions of C_(16)H_(35)NO_2,C_(18)H_(39)NO_3and C_(20)H_(43)NO_4. Active substances of C. oleifera leaf in three ecotypes weresignificant difference; each region has some different resistance-substances in leaves.Moreover, Fengyang No.4contained sheath saddle alcohol substances, such as C_(16)H_(35)NO_2and C18H39NO3of sphingosine, with the exception of fatty acyl amine substance, which isa group of significant resistance substances and significantly increased the resistance ofFengyang No.4located in the northern edge region.
1.不同生态类型区的6组油茶4个物种气孔密度的测定结果表明，江西小果、普通油茶（黄山、大别山、凤阳油茶）、博白大果与攸县油茶的气孔密度差异较大且显著，分别为6.50个/mm~2、4.35个/mm~2（普通油茶气孔密度均值）、3.20个/mm~2、2.8个/mm~2，而黄山、大别山、凤阳油茶等普通油茶同一物种间的气孔密度无显著差异。
2.通过对不同生态区域油茶品种叶片解剖结构进行观察测定发现，不同地区间油茶叶片的上下表皮、角质层、栅栏组织、海绵组织厚度等叶片表皮结构差异十分明显，且上述组织厚度由大到小排序为：凤阳油茶>大别山油茶>黄山油茶。由此可证明，凤阳地区油茶品种为适应北方低温、干旱等气候条件，叶片组织在解剖上形成了抵抗逆境环境的结构特征。
3．不同生态区域年平均降雨量与油茶叶片气孔开度和叶片细胞结构的通径分析结果表明：年平均降雨量与油茶叶片气孔长度、气孔个数、气孔开度呈正相关；但与叶片角质层的厚度却呈负相关，且有明显的直接作用。另外，在对不同生态区域油茶叶片气孔开度与叶片细胞结构中的其他各项指标多元逐步回归分析中发现，叶片气孔开度与气孔尺寸有直接正相关关系，但与叶片上、下表皮的厚度关系均为负相关。
4．本文通过DPS数据处理软件对来自三个生态区域的6个油茶物种的叶片细胞气孔开口大小进行聚类分析，结果表明：在遗传距离为1.06cm处，6个油茶种质资源共分为5个类群。类群1包括博白大果（2号）和江西小果（5号）2个物种，它们的样品相似性最大，亲缘关系最为接近。凤阳油茶亲缘关系较远，可能是该油茶长期生长在相对温度较低、降雨量较少的环境下，导致在形态上产生了较大的变异。4号攸县油茶与其他的油茶物种亲缘关系最远，其主要原因可能是物种的不同。
5．通过对不同生态区域油茶植株含水量及电导率测定发现，油茶叶片电导率以皖东地区偏低，但三个生态类型区的差异并不显著。植株新梢、叶片含水量的测定结果表明：皖南>皖西>皖东，即植株含水量与降雨量表现出明显的正相关关系；在同一生态类型区，油茶新梢含水量高于叶片含水量。
6．通过对不同生态区域油茶叶片丙二醛（MDA）及脯氨酸（Pro）含量的测定发现，三地区油茶叶片MDA含量排列顺序为：皖南地区3.227μg/g>皖西地区2.836μg/g>皖东地区2.675μg/g；当年生叶片Pro含量皖南油茶最高，达到1192.963μg/g，依次为皖西和皖东，分别为1158.938μg/g和997.129μg/g，其排序与丙二醛含量相一致。该结果表明：地处油茶分布最北缘地带的皖东地区油茶抗逆性较强。
7．通过对三个生态区油茶SOD、CAT和POD三种酶活性的测定试验结果表明：皖东地区油茶植株的SOD活性高于皖西和皖南；油茶植株CAT活性也是皖东地区高于皖南和皖西地区，但它们之间的差异不如SOD那样显著；与以上相反，油茶植株的POD活性皖东地区为最低（仅为1.5061U/g.min），而皖西和皖南地区均较高，都达到了5.2U/g.min以上。
8．通过对同一生态区域油茶品种野外调查及对其SOD、POD和CAT三种酶活性的进行了比较分析，SOD、CAT酶活性在同一地区良种的含量高低排序中出现高度的一致，分别为：皖东地区凤阳4号>凤阳3号>凤阳2号>凤阳1号>凤阳5号；皖西地区大别山3号>大别山1号>大别山4号>大别山2号；皖南地区黄山1号>皖徽5号>皖宁5号>皖宁2号>黄山2号>皖徽3号>黄山4号。这说明了SOD、CAT两种酶在油茶保护酶系统中的共同作用和核心地位。
另外多重比较分析结果表明，同一地区不同油茶良种之间POD活性差异显著，皖东凤阳2号POD活性最低，只有0.736U/g.min，凤阳1号和凤阳3号POD活性较高且差别不大；皖西仅大别山2号POD活性较低，其它良种POD活性较高，且均在6.762U/g.min以上；皖南油茶良种POD活性为黄山2号和皖宁5号含量较低，但和其他良种POD活性差异显著。
9．不同地区油茶品种的内含物研究结果表明：在所测的油茶试样中，含水量、苯醇抽提物、综纤维素、纤维素和木质素的含量分别为：44.3%-50.1%、10.8%-26.2%、0.28%-0.57%、0.12%-0.25%和0.13%-0.28%。在同一生态类型区，油茶体内仅苯-醇抽提物含量为叶片高于枝条，其余物质含量均是枝条高于叶片。在三类生态类型区中，最北缘（凤阳）油茶林的枝条、叶片苯醇抽提物含量、纤维素及木质素含量最高；含水量（枝和叶）和综纤维素（枝）为最低；其中油茶枝条的综纤维素含量差异极显著(P=0.0084<0.01）、纤维素含量差异显著(P=0.0214<0.05)；叶片综纤维素含量差异显著(P=0.0467<0.05)；由此可见，油茶体内较低的水分，较高的苯醇抽提物、纤维素和木质素含量有利于植株抗寒和抗旱能力的提高。
10．通过用高分辨质谱联用法（HPLC-DAD-MS）对皖西的大别山3号、皖南的黄山1号、皖东的凤阳4号叶片的甲醇和乙酸乙酯混合提取物进行了比较分析，发现生长在三个不同生态区域油茶品种中，大别山3号特有的化合物阴离子有C_(18)H_(24)O_(11)、C_(21)H_(20)O_(11)，阳离子有C_(23)H_(18)N_2O_4、C_(24)H_(30)O_6和C_(12)H_(20)N_4O_5；黄山1号特有的化合物有C_(27)H_(36)O_(12)；凤阳4号特有的化合物阴离子有C_(32)H_(42)O_3，阳离子有C_(16)H_(35)NO_2、C_(18_H_(39)NO_3和C_(20)H_(43)NO_4。三种生态类型的油茶叶片的活性化学物质差别较大，每个地区的叶片内都具有不同的抗性物质。其中皖东地区的凤阳4号阳离子化合物除含有脂肪酰胺类物质，还具有C_(16)H_(35)NO_2和C_(18)H_(39)NO_3等鞘氨醇类物质，鞘鞍醇类物质是一组具有显著抗性的活性物质，这也可能显著提高了处在北缘地区的凤阳4号油茶的抗性。
Camellia oleifera Abel. is woody oil tree having economic, ecological and socialbenefits, which is widely planted in recent years. Anhui province is a climate transitionregion from warm temperate zone to subtropical, which is one of C. oleifera appropriatedistribution area and is on the northern margin of C. oleifera. Thus, the factors affecting thedevelopment of C. oleifera are complicated. To promote the health development of AnhuiC. oleifera industry and to provide more theoretical basis for jianghuai region,21C.oleifera varieties from eastern, western and southern Anhui were selected as experimentalmaterials, and leaves physiological and biochemical characteristics such as Anatomystructures, enzymes activity, conductivity, benzene-ethanol extracts and cellulose content,and the differences of active substances in different ecotypes were studied, the resultsshowed that as follows:
1. Stomatal density of4species,6groups C. oleifera in different ecotypes weredetermined and the results indicated that there was significant difference between C.meiocarpa, C. oleifera (Huangshan, and dabieshan and fengyang C. oleifera), C.gigantocarpa and C.yuhsienensis, with the value of6.50a/mm~2,4.35a/mm~2(mean valueof C. oleifera stomatal density),3.20a/mm~2,2.8a/mm~2respectively. However, there wasno significant difference in stomatal density level between the same species of C. oleifera.
2. Leaf anatomical structures of C. oleifera varieties in different ecotypes weredetermined and the results showed that there was significantly different in leaves upper andlower epidermis, cutin layer, stockade and spongy tissue thickness levels. Moreover, theordering of the above tissue thickness of C. oleifera in different ecotypes was as follows:Fengyang> Dabieshan> Huangshan. Therefore, the results indicated that leaf tissuestructure of Fengyang C. oleifera formed the characteristic structure, and to adapt to thecold, drought and other adverse northern climate conditions.
3. The path analysis between and stomatal opening and leaf cell structure of C.oleifera were carried out and indicated that average annual rainfall in different ecotypeswere positively correlated with leaves stomatal length, stomatal number per unit andstomatal opening, while negatively correlated with leaf cutin layer thickness. In addition, the multiple stepwise regressions between the leaf stomatal opening and the otherindicators in leaf cell structure of C. oleifera in different ecotypes showed that leafstomatal opening was positively correlated with stomatal length, and negatively correlatedwith leaves upper and lower epidermis thickness.
4. leaf cell stomatal opening of6C. oleifera cultivars in three ecotypes were clusteranalyzed by DPS data processing software, and the results showed that6C. oleiferacultivars was divided into5groups, when the genetic distance was1.06cm. C.gigantocarpa (No.2) and C. meiocarpa (No.5) were grouped in Class1, which had amaximum similarity and a relatively close relationship. Fengyang C. oleifera was moredistant from other cultivars, because the cultivar grew long-term at a relatively lowtemperature and low rainfall, leading to a large variation of morphology. No.4C.yuhsienensis was the most distant from the other cultivars; the main reason was the cultivarfrom different species.
5. Water content and conductivity of C. oleifera leaves in different ecotypes wasdetermined and found that leaf conductivity of eastern Anhui was the lowest, while theconductivity in three ecotypes was not different; the ordering of water content wassouthern Anhui> western Anhui> eastern Anhui. Moreover, leaf water content waspositively correlated with rainfall, and water content of shoot was higher than that in leaf inthe same ecotype.
6. leaves MDA and Pro content of C. oleifera in different ecotypes weredetermined and the results showed that the ordering of MDA content was southern Anhui(3.227μg/g)> western Anhui (2.836μg/g)> eastern Anhui (2.675μg/g); leaves Procontent in southern Anhui was the highest, reaching1192.963μg/g, followed by westernAnhui and eastern Anhui, with the value of1158.938μg/g and997.129μg/g respectively,MDA content had the same tendency with Pro content in different ecotypes. The aboveresults indicated that eastern Anhui, which is located in the northern fringe of Anhui, had astrong resisting property.
7. SOD, CAT and POD activities in the three ecotypes were determined andanalyzed that SOD and CAT activities of eastern Anhui were higher than that of southern and western Anhui; there was significantly different in SOD activity level; on the contrary,POD activity in eastern Anhui was the lowest with the value of1.5061U/g.min, whereaswestern and southern Anhui was higher, reaching above5.2U/g.min.
8. SOD, POD and CAT activity in the same ecological region were compared andanalyzed. SOD, CAT enzyme activity in the same area had the same tendency: easternAnhui, Fengyang No.4> Fengyang No.3> Fengyang No.2> Fengyang No.1> FengyangNo.5; western Anhui, Dabieshan No.3> Dabieshan No.1> Dabieshan No.4> DabieshanNo.2; southern Anhui, Huangshan No.1> Wanhui No.5> Wanning No.5> Wanning No.2>Huangshan No.2> Wanhui No.3> Huangshan No.4. This explained the two enzymes ofSOD, CAT playing an important role in the protective enzyme system of C. oleifera.
In addition, multiple comparison analysis showed that POD activity was significantdifference between different varieties of C. oleifera in the same area. POD activity ofFengyang No.2in eastern Anhui was the lowest, only0.736U/g.min, and Fengyang No.1and Fengyang No.3was higher and the difference of POD activity in the two varieties waslittle; POD activity of Dabieshan No.2in western Anhui only was the lowest, POD activityin other cultivars was higher, reaching above6.762U/g.min; POD activity of HuangshanNo.2and No.5in southern Anhui was lower, there was significant difference between theabove cultivars and other cultivars.
9. Leaf inclusions of C. oleifera cultivars from the three regions were studied thatthe range of the content of water, benzene-ethanol extracts, holocellulose, cellulose content,lignin content of all the tested were44.3%-50.1%,10.8%-26.2%,0.28%-0.57%,0.12%-0.25%and0.13%-0.28%respectively. The same material content in different tissueof C. oleifera of the same ecological type was different, including the content ofbenzene-ethanol extracts of leaves was higher than that of branches, and the other materialcontents were quite the contrary.(3) The material content of branches and leaves weredifferent with ecotypes, the content of benzene-ethanol extracts, the cellulose content andlignin content of eastern Anhui were the highest, followed by western and southern Anhui;the water content of eastern Anhui was the lowest; there were significant differences of thecontent of holo-cellulose(P=0.0084<0.01), cellulose content (P=0.0214<0.05)of branches at the level of0.05, with the range of0.84%-2.14%; and there was significant difference inthe content of holo-cellulose(P=0.0467<0.05) of leaves at0.05; there was no difference inwater content and lignin content. In the future production of C. oleifera of northern edge,many different cultivation measures such as improving the content of benzene-ethanolextracts and the cellulose and lignin, reducing water content and holo-cellulose contentwere taken to improve the stress resistance of C. oleifera.
10. Methanol and mixed solvent of choroform and ethyl acetate extracts from leavesof Dabieshan No.3in western Anhui, Huangshan No.1in southern Anhui and FengyangNo.4in eastern Anhui were analyzed with HPLC-DAD-MS. In three ecotypes, DabieshanNo.3had unique negative ions of C18H24O11, C_(21)H_(20)O_(11), positive ions of C_(23)H_(18)N_2O_4,C_(24)H_(30)O_6and C_(12)H_(20)N_4O_5, Huangshan No.1has unique compounds of C_(27)H_(36)O_(12);Fengyang No.4had unique negative compounds of C_(32)H_(42)O_3, positive ions of C_(16)H_(35)NO_2,C_(18)H_(39)NO_3and C_(20)H_(43)NO_4. Active substances of C. oleifera leaf in three ecotypes weresignificant difference; each region has some different resistance-substances in leaves.Moreover, Fengyang No.4contained sheath saddle alcohol substances, such as C_(16)H_(35)NO_2and C18H39NO3of sphingosine, with the exception of fatty acyl amine substance, which isa group of significant resistance substances and significantly increased the resistance ofFengyang No.4located in the northern edge region.
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