日本红枫叶片呈色机理与增色效应的研究
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摘要
本试验以日本红枫(Acer palmatum)为试材,分别研究了日本红枫叶片色素成分分离鉴定及花色素苷的稳定性、温光互作对日本红枫叶片荧光参数和抗氧化酶活性的影响、高温强光下缓冲溶液及Fe2+溶液对日本红枫叶片光合作用及叶片内抗氧化酶活性的影响。日本红枫叶片色素成分分离鉴定及花色素苷的稳定性实验是在晴天上午9: 00~10: 00采集枝条中部叶片,根据日本红枫叶片类黄酮显色反应分析日本红枫叶片色素成分,根据不同温度、光照、酸碱度、金属离子等条件下叶绿素、花色素苷含量的变化分析其稳定性。温光互作对日本红枫叶片荧光参数和抗氧化酶活性的影响实验是将长势一致、生长健壮的日本红枫枝条放入盛有营养液的桶中,分别经过高温强光(35℃,PFD 1200μmol·m~(-2)·s~(-1))、低温强光(10℃,PFD 1200μmol·m~(-2)·s~(-1))、低温弱光(10℃,PFD 100μmol·m~(-2)·s~(-1))、高温弱光(35℃,PFD 100μmol·m~(-2)·s~(-1))处理,每组处理3次重复,测定处理过程中日本红枫叶片荧光参数及抗氧化酶活性的变化。高温强光下缓冲溶液及Fe2+溶液对日本红枫叶片光合作用及体内保护酶活性影响的实验是选择健壮植株,分别经过T1--对照(喷施蒸馏水)、T2--叶面喷施pH5.4缓冲溶液、T3--叶面喷施Fe2+ (10 mmol·L~(-1))溶液、T4--叶面喷施pH5.4缓冲溶液与Fe2+(10 mmol·L~(-1))溶液四组处理,每天傍晚喷施一次,溶液均匀喷洒于叶片正、背面,以形成可见的液珠为度。处理两天后开始取材,此后每天取材一次,连续6次。每组处理3次重复,研究四组处理对日本红枫叶片光合作用及叶片内抗氧化酶活性的影响。主要研究成果如下:
1、日本红枫叶片内的色素含有花色素苷、类胡萝卜素、黄酮醇-O-3-糖苷,可能含有黄酮醇、茶尔酮;光照越强、温度越高花色素苷的降解速度越快;pH4.5左右最有利于花色素苷的稳定,偏酸或偏碱都会加速花色素苷的降解;多种金属离子能减缓花色素苷的降解,Fe~(2+)效果最明显。
2、高温强光、高温弱光处理5 d后日本红枫叶片叶绿素含量降低29.1%、22.9%;高温强光使花色素苷含量降低6%;处理90 min后高温强光使叶片的Fo、Fm、Fv/Fm分别下降至处理前的80.5%、11%、36.9%;高温强光、高温弱光使叶片的叶黄素循环库降低9.7%、7.3%,(A+Z)/(V+A+Z)比值升高66.2%、61.9%;高温导致日本红枫叶片发生光抑制,诱导日本红枫叶片叶黄素循环启动;高温强光、低温强光、高温弱光都会影响日本红枫叶片内多数抗氧化酶活性,这三种处理使日本红枫叶片的正常生长受到一定程度的胁迫。
3、单独喷施缓冲溶液及同时喷施缓冲溶液、Fe~(2+)溶液都能使SOD、POD、CAT的酶活性增强(单独喷施缓冲溶液对SOD的影响除外),缓解高温强光对三种酶活性的抑制;单独喷施缓冲溶液、Fe~(2+)溶液及同时喷施两者都能使日本红枫叶片内可溶性糖含量增加,但对可溶性蛋白含量的影响不大;单独喷施缓冲溶液会导致日本红枫叶片内脯氨酸含量增多;除单独喷施蒸馏水外,其他三组处理不能缓解高温强光条件下日本红枫叶片叶绿素的降解,提高净光合速率,却能在一定程度上缓解花色素苷降解,以喷施缓冲溶液一组最为明显。单独喷施缓冲溶液能使PAL酶活性增强、细胞液的pH值偏低。
In order to investigate the isolation, identification of pigment and stability of anthocyanin in leaves of Acer palmatum atropurpureum, effects of temperature and light on the leaves of Acer palmatum atropurpureum, effects of pH value of soil and Fe~(2+) solution on the photosynthesis and the active of protect enzyme in Acer palmatum atropurpureum under high temperature and highlight, we used Acer palmatum atropurpureum as materials. In the isolation, identification of pigment and stability of anthocyanin in leaves of Acer palmatum atropurpureum experiment, central leaves were collected in morning of a clear sky 9:00~10:00. Analyzing the composition of pigment in leaves of Acer palmatum atropurpureum according to the color reaction of flavonoids. Analyzing stability of chlorophyll and anthocyanins according to the change of their contentses under different temperature, illumination, pH value and metal ion solution. In the effects of temperature and light on the leaves of Acer palmatum atropurpureum experiment, the wattles that healthy were put into ladles have nutrient solution. They were treated with the high temperature and strong light (35℃,PFD 1200μmol·m~(-2)·s~(-1)), the low temperature and strong light (10℃,PFD 1200μmol·m~(-2)·s~(-1)), the low temperature and poor light (10℃,PFD 100μmol·m~(-2)·s~(-1)), the high temperature and poor light (35℃,PFD 100μmol·m~(-2)·s~(-1)) with 3 repeats. The change of chlorophyll fluorescence and antioxidant enzyme activity of leaves in Acer palmatum atropurpureum were determined. In the effects of pH value of soil and Fe~(2+) solution on the photosynthesis and the active of protect enzyme in Acer palmatum atropurpureum under high temperature and highlight experiment, strong plants were selected and were divided equally into four groupsrandomly: T1---contrast (spraying distilled water ), T2---spraying buffer solution (pH5.4) to leaves, T3--spraying Fe~(2+) solution to (10 mmol·L~(-1)) leaves, T4---spraying buffer solution (pH5.4) and Fe~(2+) solution to (10 mmol·L~(-1)) leaves once a day. We come into material selection two days later and henceforth get material once a day lasting 6 days. Every level with 3 repeats, the effect of four treatments on the photosynthesis and the active of protect enzyme in Acer palmatum atropurpureum were investigated. The results showed as follows:
1. Leaves of Acer palmatum atropurpureum content anthocyanin, carotenoid, flavanols-O-3- glycoside, and may content flavanol, chalcone. The stronger light and higher temperature, the degradation of anthocyanin faster. It is helps greatly to the stability of anthocyanin that pH 4.5. Several kinds of metal ions may slowing the degradation of anthocyanin and the effect of Fe~(2+) was striking.
2. Effects of temperature and light on the leaves of Acer palmatum atropurpureum were studyed. The measurements showed that high temperature and highlight or dim light depress the content of chlorophyl by 29.1%, 22.9% respectively. High temperature and highlight depressed the content of anthocyanidin by 6%; depressed F0, Fm, Fv/Fm to 80.5%, 11%, 36.9% after 90 min. high temperature and highlight or dim light depress xanthophyll cycle pool by 9.7%, 7.3% , increase the deepoxidation extent of xanthophyll cycle(A+Z)/( V+A+Z),which was 29.5% and 24.6% higher than control. Photoinhibition and the circle of xanthin in leaves of Acer palmatum atropurpureum were induced under high temperature. Active of regment antioxidase were affected by high temperature and highlight, low temperature and highlight, high temperature and dim light. Regular growth of Acer palmatum atropurpureum was menaced to a great extent under the three treatments.
3. Effects of spraying suffer solution and iron fertilizer solution on the photosynthesis and the active of protect enzyme in Acer palmatum atropurpureum were analyzed. The results showed that: Activity of SOD, POD, CAT was increased after buffer solution was spraied or iron fertilizer solution was spraied at the same time (effect of buffer solution on activity of SOD was excepted ). Restraint of high temperature and highlight on activity of three enzymes was relieved. Content of dissoluble sugar was increased but content of dissoluble albumen have not marked change while spraying buffer solution, iron fertilizer solution or spraying buffer solution and iron fertilizer at same time. Effect of spraying buffer solution was trifling on vital movement in Acer palmatum atropurpureum leaves. Content of proline was increased as a result of spraying buffer solution. The treatments (except spraying distilled water) were unfit to slowing the degradation of chlorophyll and to up-regulated the net photosynthetic rates of Acer palmatum atropurpureum under high temperature and strong light, but were able to slowing the degradation of anthocyanin, the effect of spraying buffer solution was striking. Activity of PAL was increased and pH value of cell sap was on the low side in Acer palmatum atropurpureum leaves.
1、日本红枫叶片内的色素含有花色素苷、类胡萝卜素、黄酮醇-O-3-糖苷,可能含有黄酮醇、茶尔酮;光照越强、温度越高花色素苷的降解速度越快;pH4.5左右最有利于花色素苷的稳定,偏酸或偏碱都会加速花色素苷的降解;多种金属离子能减缓花色素苷的降解,Fe~(2+)效果最明显。
2、高温强光、高温弱光处理5 d后日本红枫叶片叶绿素含量降低29.1%、22.9%;高温强光使花色素苷含量降低6%;处理90 min后高温强光使叶片的Fo、Fm、Fv/Fm分别下降至处理前的80.5%、11%、36.9%;高温强光、高温弱光使叶片的叶黄素循环库降低9.7%、7.3%,(A+Z)/(V+A+Z)比值升高66.2%、61.9%;高温导致日本红枫叶片发生光抑制,诱导日本红枫叶片叶黄素循环启动;高温强光、低温强光、高温弱光都会影响日本红枫叶片内多数抗氧化酶活性,这三种处理使日本红枫叶片的正常生长受到一定程度的胁迫。
3、单独喷施缓冲溶液及同时喷施缓冲溶液、Fe~(2+)溶液都能使SOD、POD、CAT的酶活性增强(单独喷施缓冲溶液对SOD的影响除外),缓解高温强光对三种酶活性的抑制;单独喷施缓冲溶液、Fe~(2+)溶液及同时喷施两者都能使日本红枫叶片内可溶性糖含量增加,但对可溶性蛋白含量的影响不大;单独喷施缓冲溶液会导致日本红枫叶片内脯氨酸含量增多;除单独喷施蒸馏水外,其他三组处理不能缓解高温强光条件下日本红枫叶片叶绿素的降解,提高净光合速率,却能在一定程度上缓解花色素苷降解,以喷施缓冲溶液一组最为明显。单独喷施缓冲溶液能使PAL酶活性增强、细胞液的pH值偏低。
In order to investigate the isolation, identification of pigment and stability of anthocyanin in leaves of Acer palmatum atropurpureum, effects of temperature and light on the leaves of Acer palmatum atropurpureum, effects of pH value of soil and Fe~(2+) solution on the photosynthesis and the active of protect enzyme in Acer palmatum atropurpureum under high temperature and highlight, we used Acer palmatum atropurpureum as materials. In the isolation, identification of pigment and stability of anthocyanin in leaves of Acer palmatum atropurpureum experiment, central leaves were collected in morning of a clear sky 9:00~10:00. Analyzing the composition of pigment in leaves of Acer palmatum atropurpureum according to the color reaction of flavonoids. Analyzing stability of chlorophyll and anthocyanins according to the change of their contentses under different temperature, illumination, pH value and metal ion solution. In the effects of temperature and light on the leaves of Acer palmatum atropurpureum experiment, the wattles that healthy were put into ladles have nutrient solution. They were treated with the high temperature and strong light (35℃,PFD 1200μmol·m~(-2)·s~(-1)), the low temperature and strong light (10℃,PFD 1200μmol·m~(-2)·s~(-1)), the low temperature and poor light (10℃,PFD 100μmol·m~(-2)·s~(-1)), the high temperature and poor light (35℃,PFD 100μmol·m~(-2)·s~(-1)) with 3 repeats. The change of chlorophyll fluorescence and antioxidant enzyme activity of leaves in Acer palmatum atropurpureum were determined. In the effects of pH value of soil and Fe~(2+) solution on the photosynthesis and the active of protect enzyme in Acer palmatum atropurpureum under high temperature and highlight experiment, strong plants were selected and were divided equally into four groupsrandomly: T1---contrast (spraying distilled water ), T2---spraying buffer solution (pH5.4) to leaves, T3--spraying Fe~(2+) solution to (10 mmol·L~(-1)) leaves, T4---spraying buffer solution (pH5.4) and Fe~(2+) solution to (10 mmol·L~(-1)) leaves once a day. We come into material selection two days later and henceforth get material once a day lasting 6 days. Every level with 3 repeats, the effect of four treatments on the photosynthesis and the active of protect enzyme in Acer palmatum atropurpureum were investigated. The results showed as follows:
1. Leaves of Acer palmatum atropurpureum content anthocyanin, carotenoid, flavanols-O-3- glycoside, and may content flavanol, chalcone. The stronger light and higher temperature, the degradation of anthocyanin faster. It is helps greatly to the stability of anthocyanin that pH 4.5. Several kinds of metal ions may slowing the degradation of anthocyanin and the effect of Fe~(2+) was striking.
2. Effects of temperature and light on the leaves of Acer palmatum atropurpureum were studyed. The measurements showed that high temperature and highlight or dim light depress the content of chlorophyl by 29.1%, 22.9% respectively. High temperature and highlight depressed the content of anthocyanidin by 6%; depressed F0, Fm, Fv/Fm to 80.5%, 11%, 36.9% after 90 min. high temperature and highlight or dim light depress xanthophyll cycle pool by 9.7%, 7.3% , increase the deepoxidation extent of xanthophyll cycle(A+Z)/( V+A+Z),which was 29.5% and 24.6% higher than control. Photoinhibition and the circle of xanthin in leaves of Acer palmatum atropurpureum were induced under high temperature. Active of regment antioxidase were affected by high temperature and highlight, low temperature and highlight, high temperature and dim light. Regular growth of Acer palmatum atropurpureum was menaced to a great extent under the three treatments.
3. Effects of spraying suffer solution and iron fertilizer solution on the photosynthesis and the active of protect enzyme in Acer palmatum atropurpureum were analyzed. The results showed that: Activity of SOD, POD, CAT was increased after buffer solution was spraied or iron fertilizer solution was spraied at the same time (effect of buffer solution on activity of SOD was excepted ). Restraint of high temperature and highlight on activity of three enzymes was relieved. Content of dissoluble sugar was increased but content of dissoluble albumen have not marked change while spraying buffer solution, iron fertilizer solution or spraying buffer solution and iron fertilizer at same time. Effect of spraying buffer solution was trifling on vital movement in Acer palmatum atropurpureum leaves. Content of proline was increased as a result of spraying buffer solution. The treatments (except spraying distilled water) were unfit to slowing the degradation of chlorophyll and to up-regulated the net photosynthetic rates of Acer palmatum atropurpureum under high temperature and strong light, but were able to slowing the degradation of anthocyanin, the effect of spraying buffer solution was striking. Activity of PAL was increased and pH value of cell sap was on the low side in Acer palmatum atropurpureum leaves.
引文
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