大豆主茎木质素积累规律分析

doi:10.7668/hbnxb.20192719

摘要/Abstract

摘要：

为明确大豆节间木质素积累规律,深入理解木质素积累与大豆植株抗倒伏间的关系,进而为调控大豆植株表型和抗倒伏大豆新品种选育提供参考依据,选取具有代表性的有限生长习性品种Charleston,按照20,35株/m²的2种密度种植桶栽大豆,在2种不同密度下测定主茎的木质素含量,分析Charleston主茎第3节间的细胞伸长区(EZ)和次生细胞壁成熟区(MZ)木质素含量、在木质素合成过程中的4-香豆酸:辅酶A连接酶(4CL)、苯丙氨酸转氨酶(PAL)、肉桂酸脱氢酶(CAD)等关键酶活性以及对应基因的表达量。手工切片及分光光度计测定的结果表明,大豆主茎木质素含量由形态学上端向下端逐渐增加。低密度处理主茎中的木质素含量高于高密度。无论是高密度还是低密度处理,单一节间(茎3)木质素含量均为MZ>EZ;低密度处理茎中4CL、PAL、CAD的活性高于高密度处理。2个密度处理下茎中4CL、PAL、CAD活性变化规律较为一致,与基因表达结果相符合。大豆植株主茎中木质素含量受到木质素合成途径中的多种酶调控。木质素合成的关键酶基因多以基因家族形式存在,同一基因家族中的酶基因在同一物种植株的相同部位表达存在差异,同一基因在同一物种的不同组织和器官中的表达情况也不尽相同。

关键词: 大豆, 主茎, 木质素含量, 木质素合成关键酶

Abstract:

To clarify the lignin accumulation ways in the stem of soybean variety,and further understand the relation between the lignin accumulation and lodging resistance of soybean plant,which could give the theory evidence for plant phenotype regulation and new soybean variety breeding under different planting densities,Charleston,a representative variety with determinate growth habit,was selected in current study and were planted with two densities of 20,35 plants/m².The lignin content in the elongation zone(EZ)and the secondary cell wall maturation zone(MZ)at the third internode of Charleston's stem were measured and the activities of key enzymes such as 4-Coumarate:coenzyme A ligase(4CL)and phenylalanine aminotransferase(PAL)and cinnamate dehydrogenase(CAD)involved in the lignin synthesis were determined at two different densities.The results showed that lignin content in the stem of Charleston under low density was higher than that under high density.The content of lignin in single internode(stem 3)of Charleston under both high and low density treatments was MZ>EZ.4CL,PAL and CAD activities in low density Charleston stem were higher than those in high density of Charleston and in accordance with gene expression levels.The key genes for lignin synthesized were distributed in soybean genome in different gene families.Moreover,different enzyme gene in one gene family showed different expression style in the same section.Same results could be observed in different organ for one gene.

Key words: Soybean, Stem, Lignin content, Key enzymes for lignin synthesis

李艳燕, 赵彩桐, 齐阳阳, 刘明, 张书瑞, 刘志华, 李文滨, 姜振峰. 大豆主茎木质素积累规律分析[J]. 华北农学报, 2022, 37(3): 77-85. doi: 10.7668/hbnxb.20192719.

LI Yanyan, ZHAO Caitong, QI Yangyang, LIU Ming, ZHANG Shurui, LIU Zhihua, LI Wenbin, JIANG Zhenfeng. Analysis on the Accumulation of Lignin in the Stem of Soybean[J]. Acta Agriculturae Boreali-Sinica, 2022, 37(3): 77-85. doi: 10.7668/hbnxb.20192719.

http://www.hbnxb.net/CN/Y2022/V37/I3/77

图/表 8

表1 4CL活性测定方法

Tab.1 Method procedure for 4CL activity

试剂名称 Reagent name	测定管/μL Measuring tube
样本 Sample	160
试剂一 Reagent 1	480
试剂二 Reagent 2	80
试剂三 Reagent 3	80

图1 高密度处理下Charleston主茎间苯三酚染色 A、B、C分别为第1节茎段的上、中、下3个部位染色结果;D、E、F分别为第2节茎段的上、中、下3个部位染色结果;G、H、I分别为第3节茎段的上、中、下3个部位染色结果。图2同。

Fig.1 Resorcinol staining results of stem of Charleston under high density condition A,B and C are the staining results of the upper,middle and lower parts of the first stem segment respectively;D,E and F are the staining results of the upper,middle and lower parts of the second stem segment respectively;G,H and I are the staining results of the upper,middle and lower parts of the third stem segment,respectively.The same as Fig.2.

图2 低密度处理下Charleston主茎间苯三酚染色

Fig.2 Resorcinol staining results of stem of Charleston under low density condition

图3 高低密度处理下Charleston茎中木质素含量 A.Charleston不同节间木质素含量:CH.高密度Charleston,CL.低密度Charleston;B.茎3中EZ、MZ中木质素含量:EZ.细胞伸长区,MZ.次生细胞壁成熟区,HⅢ.高密度处理下DN594茎3,LⅢ.低密度处理下Charleston茎3。图中不同小写字母表示差异显著(Duncan's法,P<0.05)。图4-6同。

Fig.3 Lignin content of stem of Charston under high and low densities A.Charleston different internode lignin content:CH.Charleston at high density,CL.Charleston at low density;B.Lignin content of EZ and MZ in stem 3:EZ.Cell elongation region,MZ.Mature area of secondary cell wall,HⅢ.Stem 3 of Charleston under high density treatment,LⅢ.Stem 3 of Charleston under low density treatment.Different lowercase letters indicate significant difference (Duncan's multiple-range test,P<0.05). The same as Fig.4-6.

表2 高密度条件下苯丙烷合成途径代谢物含量及显著表达基因

Tab.2 Metabolite content of phenylpropane synthesis pathway and related genes under high density conditions

代谢物 Metabolites	EZ/AS	MZ/EZ	基因 Gene
肉桂酸 Cinnamic acid	-2.1		PAL	Glyma.13g145000	-3.77
				Glyma.20g180800	-2.30
				Glyma.03g181600	3.22
对香豆醇 P-coumarylalcohol	2.4		4CL	Glyma.13g372000	-2.18
松柏醛 Coniferylaldehyde	1.7	2.8	CCR	Glyma.13g369800	-0.29
				Glyma.08g218100	-1.10
松柏醇 Coniferyl alcohol	1.7	4.0	CAD	Glyma.20g128600	-1.80

图4 茎中PAL活性及不同密度处理下茎3 EZ、MZ中PAL基因表达量 A.高密度处理下Charleston第3节茎EZ、MZ中PAL基因表达量;B.高低密度处理下Charleston茎中PAL活性:Ⅱ.第2节间,Ⅲ1.第3节间EZ,Ⅲ2.第3节间MZ,Ⅳ.第4节间。图5-6同。

Fig.4 PAL activity and PAL genes in EZ and MZ of stem of Charleston in different density A.PAL gene expression levels of EZ and MZ in stem of Charleston section 3 under high density treatment;B.PAL activity in stem of Charleston treated with high and low density:Ⅱ.The second internode,Ⅲ1.The third internode EZ,Ⅲ2.The third internode MZ,Ⅳ.The fourth internode.The same as Fig.5-6.

图5 茎中4CL活性及不同密度处理下茎3 EZ、MZ中4CL基因表达量 A.高密度处理下Charleston第3节茎EZ、MZ中4CL基因表达量;B.高低密度处理下Charleston茎中4CL活性。

Fig.5 4CL activity and 4CL genes in EZ and MZ of stem of Charleston in different density A.4CL gene expression levels of EZ and MZ in stem of Charleston section 3 under high density treatment; B.4CL activity in stem of Charleston treated with high and low density.

图6 茎中CAD活性及不同密度处理下茎3 EZ、MZ中CAD基因表达量 A.高密度处理下Charleston第3节茎EZ、MZ中CAD基因表达量;B.高低密度处理下Charleston茎中CAD活性。

Fig.6 CAD activity and CAD genes in EZ and MZ of stem of Charleston in different density A.CAD gene expression levels of EZ and MZ in stem of Charleston section 3 under high density treatment; B.CAD activity in stem of Charleston treated with high and low density.

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