苹果机械伤模型与抗病生理研究
摘要
苹果的营养价值非常丰富,是人们日常食用的主要水果之一。但由于果皮较薄,在采收、分级、包装、运输、贮藏、加工和销售整个过程的各个环节中都容易受到机械损伤,从而加速营养成分的损失,加快苹果的衰老、腐败,最终导致其食用品质和商品价值的下降。
本文以苹果作为实验材料,通过实验室模拟刺伤,结合优化的台盼蓝染色方法,研究了苹果伤口周边细胞的死亡以及病菌侵染对于腐烂的影响,并对伤口组织进行分离提取,检测木质素、总多酚等抑菌物质的含量变化,结合牛津杯抑菌实验,评价果实潜在产生新细胞的能力和抗病能力；在伤口附近及远离伤口部位接种致病菌,检测腐烂大小与腐烂率,判断局部和系统抗病性的诱导；同时探讨了机械伤时间、伤液与H202对于伤口抗病能力的影响。研究结果表明：伤口细胞的死亡会逐渐影响周边细胞,使其缓慢坏死。伤口被病原菌侵染后会加速腐烂进程,而低温下发病迟缓,有利于果蔬的贮藏。刺伤部位可以诱导苹果局部和系统抗病性的产生。牛津杯抑菌实验说明抗菌成分可能是一些大分子物质,如特殊蛋白质等。伤口组织涂抹伤液后可以显著提高抗病能力,且机械伤时间越长,抗病能力越强。而利用H202诱导细胞死亡后,伤口腐烂大小与腐烂率都大大增加。
本文还通过离心实验模拟碰伤,以苹果种子幼根作为材料,研究了碰伤对细胞的形态结构及活力的影响。利用台盼蓝染色,结合生理指标的测定,试图寻找介于正常和死亡之间的伤残细胞,同时探讨了细胞死亡与液泡渗漏或细胞膜破裂的关系。实验结果表明：600r/min转速附近可能会出现伤残细胞,取两边离心速度的材料,分别检测氨基酸、脂肪酸及水杨酸的含量,差异程度较大,说明了伤残细胞存在的可能性。在800r/min转速下,细胞内出现大量气泡,细胞表面出现皱缩,可能生理异常细胞内产生气体；5000r/min转速下,中性红染液全部释放至幼根表层,细胞内没有染液滞留,可能是细胞膜发生破裂。由于缺少系统研究,相关报道也比较少,实验结论还需进一步探讨验证。
Apples with quite rich nutrition are the most commonly consumed fruits in people's daily life. Because the skin is thin, apples are prone to mechanical damage during processes such as harvesting, sorting, packaging, transportation, bulk storage and sales, which speeds up the loss of nutrients, apple's aging, corruption and directly leds to the decline of its food quality and the commodity value.
This paper reported on using the laboratory simulation stabbing apples for measurement of effects of the dead cells surrounding wound and bacterias on corruption in apple fruit, with optimization of Trypan blue staining method. Extraction and detection of lignin, total polyphenols from the wound tissues, combing with Oxford cup antibacterial experiment, was to evaluate the ability of fruit potential generating new cells and resisting diseases. Pathogenic bacteria was inoculated surrounding the wound parts and parts far away from it with detection of rotten size and rate, judging the localized and systemic induced resistance. The paper also discussed the effects of mechanical damage time, wound juice and H2O2on wound disease resistance. The results demonstrated that: the dead wound cells could gradually influence surrounding cells, making them die slowly. The wound infected by pathogenic bacteria could accelerate decay processes, while low temperature was able to delay this behavior and was helpful for storage of fruits and vegetables. Stabbed parts could induce the local and system of an apple to generate disease resistance. Oxford cup antibacterial experiments demonstrated that the antimicrobial components might be some macromolecular substances, such as special protein, etc. The wound tissues after daubing wound juice could greatly improve the disease resistance. And the longer mechanical damage time, the stronger resistance. But rotten size and rate of the wound hole increased greatly after using H2O2for inductive death on them.
This paper also studied the effect of impact on the structure formation and vigor of cells by means of running some simulations of impact with centrifugal experiments on young roots of apple seeds. Trypan Blue staining was used to find disabled cells between normal condition and death, combining with the determination of physiological indexes. The paper also discussed relations between the cell death and vacuole leakage or cell membrane rupture. The experimental results revealed that:disabled cells might appear round600rpm and there was significant difference in detection of amino acids, fatty acids and salicylic acid, taking its both sides of the centrifugal materials, which showed the possibility of existence of disabled cells. Massive bubbles appeared in cells and cell surface appeared shrinking at800rpm, which suggested that damage led to the generation of physiological abnormal cells with a mass of gas in the cells. At5000rpm, neutral red staining solution was released to the surface of young roots, without dye retention in cells and this could be due to bursting of cell membranes. Due to the lack of system research and relevant reports, the experimental conclusion still needs further discussion and validation.
本文以苹果作为实验材料,通过实验室模拟刺伤,结合优化的台盼蓝染色方法,研究了苹果伤口周边细胞的死亡以及病菌侵染对于腐烂的影响,并对伤口组织进行分离提取,检测木质素、总多酚等抑菌物质的含量变化,结合牛津杯抑菌实验,评价果实潜在产生新细胞的能力和抗病能力；在伤口附近及远离伤口部位接种致病菌,检测腐烂大小与腐烂率,判断局部和系统抗病性的诱导；同时探讨了机械伤时间、伤液与H202对于伤口抗病能力的影响。研究结果表明：伤口细胞的死亡会逐渐影响周边细胞,使其缓慢坏死。伤口被病原菌侵染后会加速腐烂进程,而低温下发病迟缓,有利于果蔬的贮藏。刺伤部位可以诱导苹果局部和系统抗病性的产生。牛津杯抑菌实验说明抗菌成分可能是一些大分子物质,如特殊蛋白质等。伤口组织涂抹伤液后可以显著提高抗病能力,且机械伤时间越长,抗病能力越强。而利用H202诱导细胞死亡后,伤口腐烂大小与腐烂率都大大增加。
本文还通过离心实验模拟碰伤,以苹果种子幼根作为材料,研究了碰伤对细胞的形态结构及活力的影响。利用台盼蓝染色,结合生理指标的测定,试图寻找介于正常和死亡之间的伤残细胞,同时探讨了细胞死亡与液泡渗漏或细胞膜破裂的关系。实验结果表明：600r/min转速附近可能会出现伤残细胞,取两边离心速度的材料,分别检测氨基酸、脂肪酸及水杨酸的含量,差异程度较大,说明了伤残细胞存在的可能性。在800r/min转速下,细胞内出现大量气泡,细胞表面出现皱缩,可能生理异常细胞内产生气体；5000r/min转速下,中性红染液全部释放至幼根表层,细胞内没有染液滞留,可能是细胞膜发生破裂。由于缺少系统研究,相关报道也比较少,实验结论还需进一步探讨验证。
Apples with quite rich nutrition are the most commonly consumed fruits in people's daily life. Because the skin is thin, apples are prone to mechanical damage during processes such as harvesting, sorting, packaging, transportation, bulk storage and sales, which speeds up the loss of nutrients, apple's aging, corruption and directly leds to the decline of its food quality and the commodity value.
This paper reported on using the laboratory simulation stabbing apples for measurement of effects of the dead cells surrounding wound and bacterias on corruption in apple fruit, with optimization of Trypan blue staining method. Extraction and detection of lignin, total polyphenols from the wound tissues, combing with Oxford cup antibacterial experiment, was to evaluate the ability of fruit potential generating new cells and resisting diseases. Pathogenic bacteria was inoculated surrounding the wound parts and parts far away from it with detection of rotten size and rate, judging the localized and systemic induced resistance. The paper also discussed the effects of mechanical damage time, wound juice and H2O2on wound disease resistance. The results demonstrated that: the dead wound cells could gradually influence surrounding cells, making them die slowly. The wound infected by pathogenic bacteria could accelerate decay processes, while low temperature was able to delay this behavior and was helpful for storage of fruits and vegetables. Stabbed parts could induce the local and system of an apple to generate disease resistance. Oxford cup antibacterial experiments demonstrated that the antimicrobial components might be some macromolecular substances, such as special protein, etc. The wound tissues after daubing wound juice could greatly improve the disease resistance. And the longer mechanical damage time, the stronger resistance. But rotten size and rate of the wound hole increased greatly after using H2O2for inductive death on them.
This paper also studied the effect of impact on the structure formation and vigor of cells by means of running some simulations of impact with centrifugal experiments on young roots of apple seeds. Trypan Blue staining was used to find disabled cells between normal condition and death, combining with the determination of physiological indexes. The paper also discussed relations between the cell death and vacuole leakage or cell membrane rupture. The experimental results revealed that:disabled cells might appear round600rpm and there was significant difference in detection of amino acids, fatty acids and salicylic acid, taking its both sides of the centrifugal materials, which showed the possibility of existence of disabled cells. Massive bubbles appeared in cells and cell surface appeared shrinking at800rpm, which suggested that damage led to the generation of physiological abnormal cells with a mass of gas in the cells. At5000rpm, neutral red staining solution was released to the surface of young roots, without dye retention in cells and this could be due to bursting of cell membranes. Due to the lack of system research and relevant reports, the experimental conclusion still needs further discussion and validation.
引文
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