Morphological Structure, Physiological And Biochemical Characteristics Of Camellia Oleifera Leaf In Different Ecotypes From Anhui Province

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/mm²,4.35a/mm²（mean valueof C. oleifera stomatal density）,3.20a/mm²,2.8a/mm²respectively. 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, C₂₁H₂₀O₁₁, positive ions of C₂₃H₁₈N₂O₄,C₂₄H₃₀O₆and C₁₂H₂₀N₄O₅, Huangshan No.1has unique compounds of C₂₇H₃₆O₁₂;Fengyang No.4had unique negative compounds of C₃₂H₄₂O₃, positive ions of C₁₆H₃₅NO₂,C₁₈H₃₉NO₃and C₂₀H₄₃NO₄. 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 C₁₆H₃₅NO₂and 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.