Ca~(2+)和钙调素拮抗剂对茄子幼苗抗冷性的影响
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摘要
茄子属典型的喜温蔬菜，低温冷害和养分缺乏是制约茄子越冬生产的两大障碍因子。钙不仅是植物必需的大量元素，而且作为偶联胞外信号与胞内生理反应的第二信使，还参与植物对外界的反应与适应，在调节植物细胞对逆境环境胁迫信号转导过程中起着重要的作用。W7作为一种新的CaM拮抗剂，可导致Ca~(2+)·CaM信使系统功能发生障碍，与CPZ(氯丙嗪)、TFP(三氟拉嗪)等其它CaM拮抗剂相比，W7具有不影响细胞膜的结构和流动性的优势。随着冬春蔬菜生产的蓬勃发展，嫁接是克服连作障碍和实现蔬菜周年供应的主要手段，但如何提高嫁接苗本身对低温胁迫的抵抗能力是可持续农业生产中的一个重要问题。目前Ca~(2+)提高蔬菜抗病性的研究较多，但关于Ca~(2+)与植物抗冷性的研究多以大田作物为材料，尚未见对嫁接苗逆境环境和矿质营养双重胁迫相互作用的报道。本试验首次通过水培供钙与缺钙处理茄子嫁接苗，W7和Ca~(2+)浸泡种子、叶片处理，从阻碍和增强Ca~(2+)·CaM信使系统转导两个角度研究钙对茄子幼苗生长和抗冷性的影响，揭示Ca~(2+)·CaM信使在植物抗冷中的作用机理，为茄子和蔬菜设施栽培防止低温冷害提供理论依据。
1．茄子嫁接苗(丰研二号／刺茄、丰研二号／野茄二号)为材料，通过水培供钙与缺钙处理，研究钙对茄子嫁接苗生长和抗冷效应。结果表明在低温胁迫下缺钙处理茄子嫁接苗和自根苗叶片保护酶(SOD、POD、CAT)活性、可溶性糖、蛋白和热稳定蛋白、游离钙和结合钙含量、UFA、U／S、IUFA值显著降低，冷害指数、电解质渗透率、MDA和SFA含量增加。与缺钙处理相比，供钙处理呈现相反的变化趋势。在相同的低温胁迫时间内，嫁接苗保护酶(SOD、POD、CAT)活性、UFA含量、U／S、IUFA显著高于自根苗。表明钙在保护茄子嫁接苗细胞膜的完整性、增加碳水化合物含量和提高植株抗冷性调控过程中起着重要的作用。
2．研究钙调素拮抗剂W7和Ca~(2+)浸种处理对茄子(丰研二号)幼苗抗冷性的影
响。结果表明W7浸种处理显著提高了低温胁迫下茄子幼苗叶片电解质渗透率和
＊OA含量，降低了SOD、＊AT活性和可溶性蛋白质含量：而k‘”处理显著降低
了低温胁迫下叶片屯解质渗透率和MDA含量，提高了SOD、CAT活性和可溶性
蛋白质含量，表明Ca’“·CaM信使系统在茄子幼苗抗冷调控过程中起着重要的作
用。
3．在低温胁迫F利用不同浓度W7和Ca’“浸叶（丰研二号）处理，结果表明：
随着W7浓度的增加，冷害后幼苗叶片的保护酶活性和可溶性糖、蛋白含量呈降
低的趋势，MDA含量增加，而随着CaZ”浓度的增加，冷胁迫后幼苗叶片内保护酶、
可溶性糖、蛋白含量丫先上升后下降的变化趋势，MDA含量先卜降后上升，并且
Ca’”在20mmol／L时浸泡叶片抗冷效果最好。
4．在不同低温胁迫时间内 W7和比”浸叶（丰研二号）处理研究 Ca‘“的抗冷效
应。试验发现，随低温时间的延长W7处理保护酶活性、可溶性糖、可溶性蛋白
含量显共降低，并显著低于对照，MDA含量显著上升并高于对照；Ca‘“处理叶片
的保护酶活性、可溶性蛋白、可溶性糖含量随低温胁迫时间的延长不先上升后卜
降的变化，并显著高于对照，而MDA含量稍有增加但仍显著低于对照。
5．本试验系统研究了茄于嫁接苗耐低温机理，并首次将钙营养和钙调素桔抗剂
应川到茄苗低温逆境研究中，结果表明钙激活了茄苗抗冷的各种指标，Ca‘“·CaM
信使系统能够稳定细胞膜的结构，使细胞膜膜脂过氧化水平降低，增加保护酶、
保护性物质和不饱和脂肪酸含量。原因可能是Ca‘”与CaM结合后调控了蛋白质合
成、某些基冈的表达和转录模式，Ca’”·CaM与W7结合后不能再与靶酶结合导
致低温胁迫后细胞膜受损，抗冷基因不能转录和表达，植株抗冷力下降。本试验
证实了 Ca‘“和 Ca’“·CC在茄茵抗冷调控中起重要作川，本研究进一步丰富了蔬
菜栽培生理学和植物嫁接植物抗冷机理。
Eggplant is a typical temperature favored vegetable. The dangerous of low temperature and nutritional-deficiency are the two main limited factors in it's winter culturing. Calcium is not only a major element of plant necessary but also a second message involved in responding and adapting to outside environment. Ca"+ plays an important role in message transducting of the environmental stress. Message transduction of calcium has showed to be inhabited by Calcium special antagonist W7. W7 can cause the barrier of Ca2* CaM message system. Not like CPZ, TFP and others antagonist, W7 doesn't affect the structure and fluidity of cell membrane. As the rapid development of greenhouse product, grafting is a main means to cope with the barrier of successive planting and realizing the vegetable supply in the whole year. But how increase the chilling- resistance of grafted seedlings is the main problem in the lasting agriculture. Recently the studies are abundant cornering to the Ca2+ increase the disease resistance of v
egetable. The study as to the Ca2+ and plant chilling resistance usually uses die field crop. There is not the study as to the relationship between environmental stresses and nutritional element of grafted seedlings.
The paper used the eggplant seedlings as the experimental materials. The grafted and own-root eggplant seedlings were cultured on solution of Calcium sufficiency and Calcium deficiency respectively to estimate the effect of Calcium and on chilling resistance of eggplant seedlings. Seeds and leaves of eggplant variety were immersed on Calcium and W7 at the different chilling time. The paper studied the effect of Calcium on growth and cold resistance of eggplant seedlings from increase and inhibiting aspects. The paper stated the theory of Ca2+-CaM chilling resistance and offered a scientific warranty to the eggplant and vegetable prevented the danger of temperature.
1. To assess the effects of calcium on growth and chilling resistance of seedlings, grafted and own-root eggplant seedlings were cultured in solution of calcium sufficiency and calcium deficiency respectively. The results showed that under calcium-deficient condition the growth of seedlings was seriously inhabited. The activities of protected enzymes and the contents of soluble sugar, soluble protein, boiling stable protein, cell inner soluble calcium and combined calcium, UFA, U/S, IUFA of calcium deficiency of grafted and own-root eggplant seedling leaves were distinctly lower than those of calcium sufficiency under low temperature condition. The activities of protected enzymes, the contents of soluble sugar, total protein, boiling stable protein, cell inner soluble calcium and combined calcium, UFA, U/S, IUFA of grafted seedlings were all obviously higher than those of own-root seedlings were all obviously higher than those
of own- root seedlings after undergoing the same time of stress. But the chilling index of electrolytic leakage, the content of MAD and SFA were direct reverse. The study suggested Ca2+ message system involved in stabling the structure of cell membranes and increasing the content of carbohydrate and enhancing cold resistance of seedlings.
2. The effect of calmodulin antagonist W7 and Ca2+ on chilling resistance of eggplant seedlings were studied. W7 treatment of immersing seeds distinctly enhanced the electrolytic leakage, the MAD content and reduced the activities of SOD, CAT and the contents of soluble sugar and protein of eggplant seedling leaves under chilling stress, but Ca2+ treatment of immersing seeds obviously decreased the contents of electrolytic leakage and MAD and increased the activities of SOD, CAT and the content of soluble sugar and protein of seedling leaves. The result suggests that Ca"+-CaM messenger system plays and important role in controlling chilling resistance of eggplant seedling.
3. The leaves of eggplant seedlings were immersed by the stable concentration of W7 and Ca** at different time. The result manifested that the activities of protected enzymes and the contents of
1．茄子嫁接苗(丰研二号／刺茄、丰研二号／野茄二号)为材料，通过水培供钙与缺钙处理，研究钙对茄子嫁接苗生长和抗冷效应。结果表明在低温胁迫下缺钙处理茄子嫁接苗和自根苗叶片保护酶(SOD、POD、CAT)活性、可溶性糖、蛋白和热稳定蛋白、游离钙和结合钙含量、UFA、U／S、IUFA值显著降低，冷害指数、电解质渗透率、MDA和SFA含量增加。与缺钙处理相比，供钙处理呈现相反的变化趋势。在相同的低温胁迫时间内，嫁接苗保护酶(SOD、POD、CAT)活性、UFA含量、U／S、IUFA显著高于自根苗。表明钙在保护茄子嫁接苗细胞膜的完整性、增加碳水化合物含量和提高植株抗冷性调控过程中起着重要的作用。
2．研究钙调素拮抗剂W7和Ca~(2+)浸种处理对茄子(丰研二号)幼苗抗冷性的影
响。结果表明W7浸种处理显著提高了低温胁迫下茄子幼苗叶片电解质渗透率和
＊OA含量，降低了SOD、＊AT活性和可溶性蛋白质含量：而k‘”处理显著降低
了低温胁迫下叶片屯解质渗透率和MDA含量，提高了SOD、CAT活性和可溶性
蛋白质含量，表明Ca’“·CaM信使系统在茄子幼苗抗冷调控过程中起着重要的作
用。
3．在低温胁迫F利用不同浓度W7和Ca’“浸叶（丰研二号）处理，结果表明：
随着W7浓度的增加，冷害后幼苗叶片的保护酶活性和可溶性糖、蛋白含量呈降
低的趋势，MDA含量增加，而随着CaZ”浓度的增加，冷胁迫后幼苗叶片内保护酶、
可溶性糖、蛋白含量丫先上升后下降的变化趋势，MDA含量先卜降后上升，并且
Ca’”在20mmol／L时浸泡叶片抗冷效果最好。
4．在不同低温胁迫时间内 W7和比”浸叶（丰研二号）处理研究 Ca‘“的抗冷效
应。试验发现，随低温时间的延长W7处理保护酶活性、可溶性糖、可溶性蛋白
含量显共降低，并显著低于对照，MDA含量显著上升并高于对照；Ca‘“处理叶片
的保护酶活性、可溶性蛋白、可溶性糖含量随低温胁迫时间的延长不先上升后卜
降的变化，并显著高于对照，而MDA含量稍有增加但仍显著低于对照。
5．本试验系统研究了茄于嫁接苗耐低温机理，并首次将钙营养和钙调素桔抗剂
应川到茄苗低温逆境研究中，结果表明钙激活了茄苗抗冷的各种指标，Ca‘“·CaM
信使系统能够稳定细胞膜的结构，使细胞膜膜脂过氧化水平降低，增加保护酶、
保护性物质和不饱和脂肪酸含量。原因可能是Ca‘”与CaM结合后调控了蛋白质合
成、某些基冈的表达和转录模式，Ca’”·CaM与W7结合后不能再与靶酶结合导
致低温胁迫后细胞膜受损，抗冷基因不能转录和表达，植株抗冷力下降。本试验
证实了 Ca‘“和 Ca’“·CC在茄茵抗冷调控中起重要作川，本研究进一步丰富了蔬
菜栽培生理学和植物嫁接植物抗冷机理。
Eggplant is a typical temperature favored vegetable. The dangerous of low temperature and nutritional-deficiency are the two main limited factors in it's winter culturing. Calcium is not only a major element of plant necessary but also a second message involved in responding and adapting to outside environment. Ca"+ plays an important role in message transducting of the environmental stress. Message transduction of calcium has showed to be inhabited by Calcium special antagonist W7. W7 can cause the barrier of Ca2* CaM message system. Not like CPZ, TFP and others antagonist, W7 doesn't affect the structure and fluidity of cell membrane. As the rapid development of greenhouse product, grafting is a main means to cope with the barrier of successive planting and realizing the vegetable supply in the whole year. But how increase the chilling- resistance of grafted seedlings is the main problem in the lasting agriculture. Recently the studies are abundant cornering to the Ca2+ increase the disease resistance of v
egetable. The study as to the Ca2+ and plant chilling resistance usually uses die field crop. There is not the study as to the relationship between environmental stresses and nutritional element of grafted seedlings.
The paper used the eggplant seedlings as the experimental materials. The grafted and own-root eggplant seedlings were cultured on solution of Calcium sufficiency and Calcium deficiency respectively to estimate the effect of Calcium and on chilling resistance of eggplant seedlings. Seeds and leaves of eggplant variety were immersed on Calcium and W7 at the different chilling time. The paper studied the effect of Calcium on growth and cold resistance of eggplant seedlings from increase and inhibiting aspects. The paper stated the theory of Ca2+-CaM chilling resistance and offered a scientific warranty to the eggplant and vegetable prevented the danger of temperature.
1. To assess the effects of calcium on growth and chilling resistance of seedlings, grafted and own-root eggplant seedlings were cultured in solution of calcium sufficiency and calcium deficiency respectively. The results showed that under calcium-deficient condition the growth of seedlings was seriously inhabited. The activities of protected enzymes and the contents of soluble sugar, soluble protein, boiling stable protein, cell inner soluble calcium and combined calcium, UFA, U/S, IUFA of calcium deficiency of grafted and own-root eggplant seedling leaves were distinctly lower than those of calcium sufficiency under low temperature condition. The activities of protected enzymes, the contents of soluble sugar, total protein, boiling stable protein, cell inner soluble calcium and combined calcium, UFA, U/S, IUFA of grafted seedlings were all obviously higher than those of own-root seedlings were all obviously higher than those
of own- root seedlings after undergoing the same time of stress. But the chilling index of electrolytic leakage, the content of MAD and SFA were direct reverse. The study suggested Ca2+ message system involved in stabling the structure of cell membranes and increasing the content of carbohydrate and enhancing cold resistance of seedlings.
2. The effect of calmodulin antagonist W7 and Ca2+ on chilling resistance of eggplant seedlings were studied. W7 treatment of immersing seeds distinctly enhanced the electrolytic leakage, the MAD content and reduced the activities of SOD, CAT and the contents of soluble sugar and protein of eggplant seedling leaves under chilling stress, but Ca2+ treatment of immersing seeds obviously decreased the contents of electrolytic leakage and MAD and increased the activities of SOD, CAT and the content of soluble sugar and protein of seedling leaves. The result suggests that Ca"+-CaM messenger system plays and important role in controlling chilling resistance of eggplant seedling.
3. The leaves of eggplant seedlings were immersed by the stable concentration of W7 and Ca** at different time. The result manifested that the activities of protected enzymes and the contents of
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