铅锌胁迫下蓖麻叶糖代谢规律及相关酶基因差异表达分析
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摘要
以蓖麻为材料,在现蕾期前持续铅锌胁迫,分析此过程中植株叶片可溶性糖和淀粉含量变化及糖代谢关键酶基因表达特点。结果表明:叶片可溶性糖含量在0.0015 mol·kg~(-1)单一Pb~(2+)胁迫处理植株中最高,其次是0.0005 mol·kg~(-1) Pb~(2+)+0.0015 mol·kg~(-1) Zn~(2+)复合胁迫处理植株;叶片淀粉含量在0.0030 mol·kg~(-1)单一Zn~(2+)处理植株中最高,其次是浓度0.0015 mol·kg~(-1)单一Pb~(2+)胁迫处理的植株。铅锌复合胁迫处理植株叶片中可溶性糖和淀粉累积量均高于对照,且当铅锌单一胁迫浓度分别为0.0020和0.0060 mol·kg~(-1)时,可溶性糖含量较对照无显著差异,淀粉含量较对照存在显著差异,同时,差异基因在淀粉和蔗糖代谢途径、糖酵解/糖异生途径中富集最多。铅胁迫下,蔗糖磷酸合成酶(SPS)、β-呋喃果糖苷酶(INV)、β-淀粉酶、果糖~(-1),6-二磷酸酶I类(FBP)、6-磷酸果糖激酶~(-1) (PFK~(-1))、磷酸丙糖异构酶(TPI)等关键酶基因表达水平发生变化;锌胁迫下1,4-α-葡聚糖分支酶(GBE1)、蔗糖合成酶(SS)、颗粒结合淀粉合成酶(WAXY)、α-与β-淀粉酶、果糖~(-1),6-二磷酸酶I类(FBP)、葡萄糖磷酸变位酶(PGM)、己糖激酶(HK)、3-磷酸甘油醛脱氢酶(GAPDH)、丙酮酸脱羧酶(PDC)、丙酮酸激酶(PK)等关键酶基因表达水平发生变化。即铅、锌胁迫下糖代谢基因差异表达,且此时可溶性糖含量较对照无显著差异,淀粉含量较对照存在显著差异,这也说明重金属铅锌胁迫影响糖代谢平衡使得植株生长期延长。
The lead and zinc stress of castor was carried out before the bud stage. The changes of soluble sugar and starch content and the expression of key enzymes gene in carbon metabolism were analyzed. The results showed that the leaf soluble content was the highest in 0.0015 mol·kg~(-1) single Pb~(2+) stress treatment, followed by 0.0005 mol·kg~(-1) Pb~(2+)+0.0015 mol·kg~(-1) Zn~(2+) combined stress treatment; the leaf starch content was the highest in 0.0030 mol·kg~(-1) among the Zn~(2+) treated plants, followed by the plants treated with a single Pb~(2+) stress at a concentration of 0.0015 mol·kg~(-1). The accumulation of soluble sugar and starch in the leaves of Pb-Zn combined stress treatment was higher than that of the control. When the single stress concentration of lead and zinc was 0.0020 and 0.0060 mol·kg~(-1), respectively, the soluble sugar content was not significantly different from the control, but the starch content had significant differences. At the same time, differential genes were most abundant in starch and sucrose metabolic pathways, and glycolysis/gluconeogenesis pathways. Under lead stress, the expression levels of genes encoding sucrose phosphate synthase(SPS), β-fructofuranosidase(INV), β-amylase, fructose~(-1),6-bisphosphatase I(FBP), 6-phosphate fructokinase~(-1)(PFK~(-1)), and triose phosphate isomerase(TPI) were changed; while under zinc stress expression levels of key genes such as 1,4-α-glucan branching enzyme(GBE1), sucrose synthase(SS), granule-bound starch synthase(WAXY), α-and β-amylase, FBP, phosphoglucomutase(PGM), hexokinase(HK), glyceraldehyde-3-phosphate dehydrogenase(GAPDH), pyruvate decarboxylase(PDC), and pyruvate kinase(PK) were changed. The expression of sugar metabolism genes was differentially expressed under lead and zinc stress; and compared with control, the starch content was significantly different, while there was no evident difference in soluble sugar content. This result also indicates that heavy metal lead and zinc stress may affect the balance of sugar metabolism and prolongs the growth period of the plant.
The lead and zinc stress of castor was carried out before the bud stage. The changes of soluble sugar and starch content and the expression of key enzymes gene in carbon metabolism were analyzed. The results showed that the leaf soluble content was the highest in 0.0015 mol·kg~(-1) single Pb~(2+) stress treatment, followed by 0.0005 mol·kg~(-1) Pb~(2+)+0.0015 mol·kg~(-1) Zn~(2+) combined stress treatment; the leaf starch content was the highest in 0.0030 mol·kg~(-1) among the Zn~(2+) treated plants, followed by the plants treated with a single Pb~(2+) stress at a concentration of 0.0015 mol·kg~(-1). The accumulation of soluble sugar and starch in the leaves of Pb-Zn combined stress treatment was higher than that of the control. When the single stress concentration of lead and zinc was 0.0020 and 0.0060 mol·kg~(-1), respectively, the soluble sugar content was not significantly different from the control, but the starch content had significant differences. At the same time, differential genes were most abundant in starch and sucrose metabolic pathways, and glycolysis/gluconeogenesis pathways. Under lead stress, the expression levels of genes encoding sucrose phosphate synthase(SPS), β-fructofuranosidase(INV), β-amylase, fructose~(-1),6-bisphosphatase I(FBP), 6-phosphate fructokinase~(-1)(PFK~(-1)), and triose phosphate isomerase(TPI) were changed; while under zinc stress expression levels of key genes such as 1,4-α-glucan branching enzyme(GBE1), sucrose synthase(SS), granule-bound starch synthase(WAXY), α-and β-amylase, FBP, phosphoglucomutase(PGM), hexokinase(HK), glyceraldehyde-3-phosphate dehydrogenase(GAPDH), pyruvate decarboxylase(PDC), and pyruvate kinase(PK) were changed. The expression of sugar metabolism genes was differentially expressed under lead and zinc stress; and compared with control, the starch content was significantly different, while there was no evident difference in soluble sugar content. This result also indicates that heavy metal lead and zinc stress may affect the balance of sugar metabolism and prolongs the growth period of the plant.
引文
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Rogers EE,Eide DJ,Guerinot ML(2000).Altered selectivity in an Arabidopsis metal transporter.Proc Natl Acad Sci USA,97:12356-12360
Sha CY,Meng QR,Wang J,et al(2011).Physiological responses of two colored-leaf species to Pb stress.J Northwest For Univ,26:36-40(in Chinese with English abstract)[沙翠芸,孟庆瑞,王静等(2011).两种彩叶植物对铅胁迫的生理响应.西北林学院学报,26:36-40]
Sun ZJ,LüLY,Wu YP(2012).Castor industry development:from cultivation to product exploitation.J China Agric Univ,17:204-214(in Chinese with English abstract)[孙振钧,吕丽媛,伍玉鹏(2012).蓖麻产业发展:从种植到利用.中国农业大学学报,17:204-214]
Yang Y,Lou YH,Yang ZJ,et al(2016).Effect of low temperature on phytohormones and carbohydrates metabolism in Bermuda grass.Acta Pratac Sin,25:205-215(in Chinese with English abstract)[杨勇,娄燕宏,杨知建等(2016).低温胁迫对狗牙根激素和碳水化合物代谢的影响.草业学报,25:205-215]
Yao G,Gao HY,Wang WW,et al(2009).The effects of Pbstress on functions of photosystems and photosynthetic rate in maize seedling leaves.Acta Ecol Sin,29:1162-1169(in Chinese with English abstract)[姚广,高辉远,王未未等(2009).铅胁迫对玉米幼苗叶片光系统功能及光合作用的影响.生态学报,29:1162-1169]
Zeng XL(2010).Research progress for resistance and tolerance mechanism of castor.Chin Agric Sci Bull,26:123-125(in Chinese with English abstract)[曾小龙(2010).蓖麻在逆境胁迫下的抗性及耐性机制研究进展.中国农学通报,26:123-125]
Zhu YR,Liu MM,Li YH,et al(2013).Research advance in regulation mechanism of starch synthesis in plants.Plant Physiol J,49:1319-1325(in Chinese with English abstract)[朱晔荣,刘苗苗,李亚辉等(2013).植物淀粉生物合成调节机制的研究进展.植物生理学报,49:1319-1325]
Chen JX,Wang XF(2006).Experimental Guidance of Plant Physiology.2nd ed.Guangzhou:South China University of Technology Press,55-56(in Chinese)[陈建勋,王晓峰(2006).植物生理学实验指导.第2版.广州:华南理工大学出版社,55-56]
Chen YH,Liu XY,Wang MX,et al(2014).Cadmium tolerance,accumulation and relationship with Cd subcellular distribution in Ricinus communis L.Acta Sci Circum,34:2440-2446(in Chinese with English abstract)[陈亚慧,刘晓宇,王明新等(2014).蓖麻对镉的耐性,积累及与镉亚细胞分布的关系.环境科学学报,34:2440-2446]
Dong JQ,Lei QS,Zhang HZ,et al(2018).Effects evaluation of phosphate stabilizes zinc contaminated soil in laboratory test and field remediation construction.Chin J Environ Eng,12:923-930(in Chinese with English abstract)[董璟琦,雷秋霜,张红振等(2018).磷酸盐稳定化修复锌污染土壤小试和工程效果评估.环境工程学报,12:923-930]
Du X,Shen XH(2010).Effects of cadmium stress on physiological and biochemical indices of Viburnum odoratissimum and V.tinus seedlings.Chin J Ecol,29:899-904(in Chinese with English abstract)[杜晓,申晓辉(2010).镉胁迫对珊瑚树和地中海荚蒾生理生化指标的影响.生态学杂志,29:899-904]
He YF,Li X,Xie YF(2016).Research progress in sugar signal and its regulation of stress in plants.Plant Physiol J,52(3):241-249(in Chinese with English abstract)[何亚飞,李霞,谢寅峰(2016).植物中糖信号及其对逆境调控的研究进展.植物生理学报,52(3):241-249]
Hou JF(2012).Sucrose content in fresh soybean seeds and cloning and functional analysis of genes related to sugar metabolism(dissertation).Nanjing:Nanjing Agricultural University(in Chinese with English abstract)[侯金锋(2012).大豆鲜籽粒蔗糖含量的研究及糖代谢相关基因的克隆与功能分析(学位论文).南京:南京农业大学]
Hu PJ,Li Z,Zhong DX,et al(2014).Research progress on the phytoextraction of heavy metal contaminated soils in China.Plant Physiol J,50:577-584(in Chinese with English abstract)[胡鹏杰,李柱,钟道旭等(2014).我国土壤重金属污染植物吸取修复研究进展.植物生理学报,50:577-584]
Rogers EE,Eide DJ,Guerinot ML(2000).Altered selectivity in an Arabidopsis metal transporter.Proc Natl Acad Sci USA,97:12356-12360
Sha CY,Meng QR,Wang J,et al(2011).Physiological responses of two colored-leaf species to Pb stress.J Northwest For Univ,26:36-40(in Chinese with English abstract)[沙翠芸,孟庆瑞,王静等(2011).两种彩叶植物对铅胁迫的生理响应.西北林学院学报,26:36-40]
Sun ZJ,LüLY,Wu YP(2012).Castor industry development:from cultivation to product exploitation.J China Agric Univ,17:204-214(in Chinese with English abstract)[孙振钧,吕丽媛,伍玉鹏(2012).蓖麻产业发展:从种植到利用.中国农业大学学报,17:204-214]
Yang Y,Lou YH,Yang ZJ,et al(2016).Effect of low temperature on phytohormones and carbohydrates metabolism in Bermuda grass.Acta Pratac Sin,25:205-215(in Chinese with English abstract)[杨勇,娄燕宏,杨知建等(2016).低温胁迫对狗牙根激素和碳水化合物代谢的影响.草业学报,25:205-215]
Yao G,Gao HY,Wang WW,et al(2009).The effects of Pbstress on functions of photosystems and photosynthetic rate in maize seedling leaves.Acta Ecol Sin,29:1162-1169(in Chinese with English abstract)[姚广,高辉远,王未未等(2009).铅胁迫对玉米幼苗叶片光系统功能及光合作用的影响.生态学报,29:1162-1169]
Zeng XL(2010).Research progress for resistance and tolerance mechanism of castor.Chin Agric Sci Bull,26:123-125(in Chinese with English abstract)[曾小龙(2010).蓖麻在逆境胁迫下的抗性及耐性机制研究进展.中国农学通报,26:123-125]
Zhu YR,Liu MM,Li YH,et al(2013).Research advance in regulation mechanism of starch synthesis in plants.Plant Physiol J,49:1319-1325(in Chinese with English abstract)[朱晔荣,刘苗苗,李亚辉等(2013).植物淀粉生物合成调节机制的研究进展.植物生理学报,49:1319-1325]