外源亚精胺对高温胁迫下生菜氮代谢的影响

doi:10.3864/j.issn.0578-1752.2022.07.011

Abstract

Abstract:

【Objective】 The lettuce yield and nutritional quality planted in high-temperature seasons are not good. Nitrogen is the most closely related nutrient element to lettuce yield. In this study, the regulation mechanism of exogenous spermidine (Spd) on nitrogen metabolism of lettuce under high-temperature stress was studied, and the changes of amino acid nutritional quality were observed, so as to provide some protective measures to reduce the harm of high temperature on lettuce. 【Method】 The test material was leaf lettuce Beisansheng 3. The test treatments were set as normal temperature (day/night: 22℃/17℃) control (CK), high-temperature stress (day/night: 35℃/30℃) treatment (H), and high temperature spraying Spd treatment (HS). The changes of lettuce morphology were observed, and the changes of the lettuce root ion flow rate, nitrogen compounds contents (total nitrogen, ammonia nitrogen, and nitrate nitrogen), and the activities of key enzymes in nitrogen metabolism as well as their relative expression levels were analyzed. In addition, the effects of the CK, H, and HS treatments on the lettuce leaf free amino acids content were studied. 【Result】 Compared with the CK, the lettuce growth of the H treatment was inhibited. Compared with the H treatment, the HS treatment increased total fresh weight, total dry weight, leaf length, leaf width, root length, root volume, and root surface area of lettuce by 24.00%, 24.62%, 14.97%, 11.83%, 23.24%, 29.47%, and 36.98%, respectively. The changes in the total nitrogen content of lettuce treated by CK, H, and HS were consistent with the growing trend of lettuce. The net absorption amount of NO₃^- in lettuce roots treated with CK, H and HS were much smaller than that of NH₄⁺, indicating that lettuce roots mainly absorbed NH₄⁺. The H treatment resulted in the outflow of NO₃^- from roots, while the HS treatment promoted the influx of NO₃^- from roots. The accumulation of ammonia nitrogen content in leaves under the HS treatment was effectively inhibited. Meanwhile, the activities of nitrate reductase (NR), glutamine synthase (GS), glutamate synthase (GOGAT), and glutamate dehydrogenase (GDH) under the HS treatment were increased and the relative gene expression levels of LsNR, LsGS, LsGOGAT1, and LsGDH in leaves and roots were up-regulated. Compared with the CK treatment, the H treatment reduced the total free amino acid (TAA) content (P<0.05); the HS treatment alleviated the reduction in TAA content caused by high temperature. The changes of umami amino acids (UAA), sweet amino acids (SAA), and bitter amino acids (BAA) among the CK, H, and HS treatment were consistent with those of TAA, while aromatic amino acids (AAA) were the opposite. The changes of aspartic acid (Asp), threonine (Thr), glutamic acid (Glu), glycine (Gly), valine (Val), lysine (Lys), histidine (His), arginine (Arg), and proline (Pro) among the CK, H, and HS treatment were consistent with those of TAA. Among the free amino acids tested, Arg had the largest coefficient of variation. 【Conclusion】Spraying Spd treatment inhibited the decrease of lettuce biomass accumulation under high temperature. For spraying Spd treatment, the nitrogen metabolism disorders under high-temperature stress was alleviated, and the leaf total free amino acid content, flavor amino acids of umami amino acids, sweet amino acids, and bitter amino acids were increased, while the high-temperature resistant capacity of lettuce was effectively enhanced and the nutritional quality of lettuce was improved.

Key words: high temperature, Lactuca sativa L., nitrogen metabolism, spermidine, ion flow rate, amino acid

YU QiLong,HAN YingYan,HAO JingHong,QIN XiaoXiao,LIU ChaoJie,FAN ShuangXi. Effect of Exogenous Spermidine on Nitrogen Metabolism of Lettuce Under High-Temperature Stress[J].Scientia Agricultura Sinica, 2022, 55(7): 1399-1410.

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References 42

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基因名称Gene name	上游引物 Forward primer (5′-3′)	下游引物 Reverse primer (5′-3′)
LsNR	CTCCGAGCCACCTTTGACTAAGC	CGCATATCTCCACCGTCCAATCC
LsGS	CTTCGCCACGCAGACCACATC	ATGGAGCAACCACGGTTAGCAAC
LsGOGAT	GCAGTTCGTGAGAAGCGTGAGAG	TGAAGCAGCGGATTCCACCTTTC
LsGOGAT1	CGTGAGGAAGATGTGCTGAAGGAG	GATGGTCGGGTTGCCTGTTCG
LsGDH	CCGTATGGTGGTGCGAAAGGTG	GAGCAGGAACATCGGTGTGGATTC
18S	GTGAGTGAAGAAGGGCAATG	CACTTTCAACCCGATTCACC

处理Treatment	总鲜重 Total fresh weight (g/plant)	总干重 Total dry weight (g/plant)	叶长 Leaf length (cm)	叶宽 Leaf width (cm)	根长 Root length (cm)	根系体积 Root volume (cm³)	根系表面积 Root surface area (cm²)
CK	15.60±0.49a	0.65±0.04b	13.78±0.25c	11.08±0.46a	24.30±0.98a	5.75±0.16a	281.96±10.49a
H	10.96±0.38c	0.65±0.73b	14.90±0.31b	7.86±0.24c	18.50±0.05c	3.19±0.35c	169.06±15.86c
HS	13.59±0.38b	0.81±0.11a	17.13±0.40a	8.79±0.28b	22.80±2.25b	4.13±0.40b	231.57±13.52b

游离氨基酸组成 Free amino acid composition (μg·g^-1)	处理 Treatment			含量均值 Average content (μg·g^-1)	变异系数 CV (%)
游离氨基酸组成 Free amino acid composition (μg·g^-1)	CK	H	HS	含量均值 Average content (μg·g^-1)	变异系数 CV (%)
天冬氨酸Asp	177.80±15.18a	100.46±10.14b	187.29±6.02a	155.18	30.69
苏氨酸Thr	528.62±6.69a	221.80±51.22c	332.73±24.36b	361.05	43.03
丝氨酸Ser	263.18±2.87a	195.94±5.54b	159.78±0.89c	206.30	25.44
谷氨酸Glu	949.11±46.19a	739.63±14.82c	839.33±32.91b	842.69	12.43
甘氨酸Gly	6.53±0.03a	4.81±0.47b	6.31±0.51a	5.88	15.91
丙氨酸Ala	128.55±1.59a	104.18±7.89b	85.38±0.93c	106.04	20.41
胱氨酸Cys	6.25±0.20a	6.39±0.16a	6.33±0.06a	6.32	1.11
缬氨酸Val	88.98±0.62b	68.14±3.20c	96.66±2.10a	84.59	17.45
蛋氨酸Met	2.90±0.04a	2.69±0.13a	2.88±0.13a	2.82	4.11
异亮氨酸Ile	17.32±0.05a	15.89±0.35b	15.41±0.6b	16.21	6.13
亮氨酸Leu	10.79±0.47a	6.77±0.22b	6.64±0.12b	8.07	29.25
酪氨酸Tyr	14.13±0.12b	16.28±0.52a	13.30±0.33c	14.57	10.56
苯丙氨酸Phe	23.63±0.08b	24.52±1.10ab	25.02±0.11a	24.39	2.89
赖氨酸Lys	14.10±0.24a	12.33±0.03b	11.55±0.36c	12.66	10.32
组氨酸His	9.50±0.21b	6.81±0.59c	12.60±1.76a	9.64	30.07
精氨酸Arg	60.55±0.71b	42.28±7.97c	101.88±7.04a	68.24	44.75
脯氨酸Pro	1085.58±30.69a	899.09±8.30c	943.91±4.74b	976.19	9.97
总氨基酸TAA	3387.54±44.14a	2468.01±63.91c	2847.01±37.26b	2900.85	15.93

呈味氨基酸组成 Flavor amino acid composition (μg·g^-1)	处理 Treatment			含量均值 Average content (μg·g^-1)	变异系数 CV (%)
呈味氨基酸组成 Flavor amino acid composition (μg·g^-1)	CK	H	HS	含量均值 Average content (μg·g^-1)	变异系数 CV (%)
鲜味氨基酸UAA	1276.09±34.67a	961.42±9.65c	1129.86±35.01b	1122.46	14.03
甜味氨基酸SAA	1886.89±31.66a	1323.64±55.74c	1449.03±28.59b	1553.19	19.04
芳香族氨基酸AAA	44.01±0.18b	47.19±1.56a	44.66±0.49b	45.29	3.71
苦味氨基酸BAA	180.54±1.36b	135.76±10.90c	223.47±9.43a	179.92	24.38

Effect of Exogenous Spermidine on Nitrogen Metabolism of Lettuce Under High-Temperature Stress

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