Impact of Acid Stress on Physiological Metabolism and Antioxidant Regulatory Gene Expression in Hainan Green Sweet Orange

doi:10.7668/hbnxb.20195886

Abstract

Abstract:

This study aimed to investigate the effects of acidic soil on the growth metabolism and photosynthesis of Hainan green sweet orange (Citrus sinensis L.)and to elucidate its antioxidant regulatory mechanisms under acid stress.A pot experiment was conducted to analyze the physiological metabolic changes,antioxidant responses,and expression patterns of key antioxidant enzyme regulatory genes in green sweet orange under acid stress.The results showed that the plant height growth,leaf area and fresh weight of green sweet orange plants decreased significantly under a strongly acidic soil environment,with the most serious effect of pH 3.5 treatment;the weakly acidic(pH 6.5)soil could promote the growth of plants,which manifested as positive growth in the amount of each growth index.Strong acid treatment significantly impaired photosynthesis and normal growth metabolism of green sweet orange leaves,manifested as negative growth in transpiration rate(Tr)and stomatal conductance(Gs)were significantly reduced.Superoxide dismutase(SOD)and phenylalanine ammonia-lyase(PAL)activities increased by 6.60-fold and 5.90-fold at 24 h post-treatment under pH 3.5.The malondialdehyde(MDA)content in leaves treated with pH 3.5 increased by 6.47-fold compared to the initial time point(0 h),significantly higher than that under pH 6.5.Principal component analysis(PCA)and redundancy analysis(RDA)revealed a significant negative correlation between enzyme activity and physiological-biochemical indices(R²=0.82).Gene expression analysis demonstrated that CsSOD and CsPAL reached peak levels at 24,48 h post-stress respectively and remained upregulated within 72 h,showing 2.33-fold and 2.64-fold increases compared to 0 h,respectively.In summary,strongly acidic soil inhibits the growth and development of green sweet orange by reducing photosynthetic pigment content and metabolic efficiency.SOD and PAL serve as key antioxidant enzymes in response to acid stress,with CsSOD and CsPAL genes dynamically regulating enzymatic activity to mitigate stress.

Key words: Green sweet orange, Acid stress, Physiological metabolism, Photosynthetic metabolism, Antioxidant enzyme, Regulatory gene

CLC Number:

ZHAO Ya, FAN Hongyan, LI Shaoka, YAN Caibin, BAI Li, HUANG Haijie, XIAO Min. Impact of Acid Stress on Physiological Metabolism and Antioxidant Regulatory Gene Expression in Hainan Green Sweet Orange[J]. Acta Agriculturae Boreali-Sinica, 2026, 41(1): 86-96. doi: 10.7668/hbnxb.20195886.

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References 53

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基因名称 Genes	引物名称 Primers	引物序列(5'—3') Primer sequences(5'—3')	产物大小/bp Product size	基因序列号 Gene ID
CsPOD	POD-F	GGAAGAGAGTCGGTTGCCT	198	Cs_ont_2g019550.1
	POD-R	TGTGTCGCCTTTGCCAGT	198
CsCAT	CAT-F	CTTTGACCCACTTGATGTAACC	149	Cs_ont_3g001020.1
	CAT-R	TGCCAGGAACCACAATAGC	149
CsSOD	SOD-F	ACAGCACAATGGTGAAAGCA	202	Cs_ont_3g017770.2
	SOD-R	GGGTTAAAGTGGGGTCCG	202
CsPAL	PAL-F	AACTCGGTGAATGACAATCCC	120	Cs_ont_6g020620.1
	PAL-R	TGCCGCAATAGCCAAACG	120
Actin	Actin-F	CCGAGCGTGGTTATTCTTTC	123	—
	Actin-R	CTGAGCTGGTCTTTGCTGTCT	123

基因名称 Genes	引物名称 Primers	引物序列(5'—3') Primer sequences(5'—3')	产物大小/bp Product size	基因序列号 Gene ID
CsPOD	POD-F	GGAAGAGAGTCGGTTGCCT	198	Cs_ont_2g019550.1
	POD-R	TGTGTCGCCTTTGCCAGT	198
CsCAT	CAT-F	CTTTGACCCACTTGATGTAACC	149	Cs_ont_3g001020.1
	CAT-R	TGCCAGGAACCACAATAGC	149
CsSOD	SOD-F	ACAGCACAATGGTGAAAGCA	202	Cs_ont_3g017770.2
	SOD-R	GGGTTAAAGTGGGGTCCG	202
CsPAL	PAL-F	AACTCGGTGAATGACAATCCC	120	Cs_ont_6g020620.1
	PAL-R	TGCCGCAATAGCCAAACG	120
Actin	Actin-F	CCGAGCGTGGTTATTCTTTC	123	—
	Actin-R	CTGAGCTGGTCTTTGCTGTCT	123

处理 Treatments	株高生长量/mm Plant height increment	叶面积/cm² Leaf area	鲜质量/g Fresh weight
pH 6.5	49.21±2.56a	2 638.86±629.01a	98.33±8.33a
pH 5.5	44.76±3.43ab	1 482.69±178.58b	78.89±9.99abc
pH 4.5	39.43±2.33b	1 284.07±79.87b	68.89±9.99bc
pH 3.5	38.38±2.80b	938.28±57.71b	59.22±5.68c

处理 Treatments	株高生长量/mm Plant height increment	叶面积/cm² Leaf area	鲜质量/g Fresh weight
pH 6.5	49.21±2.56a	2 638.86±629.01a	98.33±8.33a
pH 5.5	44.76±3.43ab	1 482.69±178.58b	78.89±9.99abc
pH 4.5	39.43±2.33b	1 284.07±79.87b	68.89±9.99bc
pH 3.5	38.38±2.80b	938.28±57.71b	59.22±5.68c

处理 Treatments	净光合速率/(μmol/ (m²·s)) Assimilation rate			蒸腾速率/(mmol/(m²·s)) Transportation rate				气孔导度/(mmol/(m²·s)) Stomatal conductance		气孔限制值 Stomatal limitation
处理 Treatments	0 h	56 d		0 h		56 d		0 h	56 d	0 h		56 d
pH 6.5	5.43±2.51a	5.45±0.39a		4.33±0.29ab		3.60±0.49a		241.67±45.95a	218.50±50.43a	-0.14			-0.12
pH 5.5	7.47±0.92a	5.87±0.40a		5.60±0.40a		2.78±0.15ab		261.00±32.71a	124.56±10.31b	-0.13			-0.23
pH 4.5	7.35±0.66a	5.40±0.66a		4.00±0.52b		2.37±0.32b		214.50±47.63a	104.75±18.05b	-0.20			-0.31
pH 3.5	6.85±0.65a	5.67±0.62a		4.73±0.35ab		3.35±0.58ab		280.29±44.28a	152.00±8.00ab	-0.14			-0.32
处理 Treatments	水分利用率/% Water use efficiency				蒸气压亏/mb Vapor pressure deficit				胞间CO₂浓度/(cm³/m³) Intercellular CO₂ concentration
处理 Treatments	0 h		56 d		0 h		56 d		0 h		56 d
pH 6.5	1.30±0.62a		1.58±0.09c		2.07±0.23a		1.93±0.17b		300.67±27.42a		298.75±4.77a
pH 5.5	1.43±0.28a		2.17±0.19b		1.65±0.19a		2.39±0.08a		303.83±14.83a		260.44±10.01b
pH 4.5	1.85±0.87a		2.63±0.22ab		2.20±0.17a		2.53±0.09a		279.00±7.51a		234.33±10.17b
pH 3.5	1.30±0.26b		2.24±0.15a		2.09±0.14a		2.08±0.07b		300.00±13.85a		229.00±8.32b

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