干旱胁迫下2种内生真菌对烟草生理生化指标及NAC基因表达的影响

doi:10.7668/hbnxb.20194526

摘要/Abstract

摘要：

为探究印度梨形孢联合丛枝菌根真菌(AMF)对烟草抗旱性的影响,在温室条件下开展盆栽试验,以云烟87为材料,设立接种无菌水(CC)、印度梨形孢(CP)、AMF(PC)、印度梨形孢和AMF(PP)处理,以自然干旱的方式进行胁迫,测定烟草叶片中脯氨酸(Pro)、超氧化物歧化酶(SOD)、过氧化物酶(POD)、丙二醛(MDA)生理生化指标和干旱相关基因NTNAC1和NAC4表达情况。结果表明,印度梨形孢和AMF均能促进烟草地上部分和地下部分生长,且干旱胁迫后褪绿不明显,干旱症状轻微。干旱胁迫7 d后,与CC组相比,CP、PC和PP组烟草叶片中Pro含量显著升高,分别为CC组的1.39,1.59,1.78倍;烟草叶片中SOD和POD活性呈现先升高后降低的趋势,CP、PC和PP组的SOD活性分别是CC组的1.15,1.22,1.33倍,POD活性分别是CC组的1.33,1.46,1.85倍;烟草叶片中MDA含量降低,CP、PC和PP组的MDA含量比CC组分别减少21.98%,23.98%,24.84%;烟草叶片干旱相关基因NTNAC1和NAC4表达上调,CP、PC和PP组NTNAC1基因表达量分别是CC组的3.37,3.88,5.07倍,NAC4基因表达量分别是CC组的3.04,3.59,5.56倍。该研究表明,印度梨形孢和AMF表现出显著的协同作用,能够显著提高烟草的抗旱性。

关键词: 烟草, 印度梨形孢, 丛枝菌根真菌, 抗旱性

Abstract:

To investigate the effects of Piriformospora indica combined with Arbuscular mycorrhizal fungi (AMF) on drought resistance of tobacco,Yunyan 87 was inoculated with sterile water (CC),P.indica (CP),AMF (PC),P.indica and AMF (PP),and the contents of proline (Pro),superoxide dismutase (SOD),peroxidase (POD),malondialdehyde (MDA) and the expression of drought-related genes NTNAC1 and NAC4 in tobacco leaves were determined under stress in the form of natural drought.The results showed that P.indica and AMF could promote the growth of above-ground and underground parts of tobacco,and chlorosis of leaves and symptoms of their drought were slight.After 7 days of drought stress,the contents of Pro in tobacco leaves of CP,PC and PP groups were 1.39,1.59 and 1.78 times higher than those of CC in group,respectively.The activities of SOD and POD in tobacco leaves increased first and then decreased.The activities of SOD in CP,PC and PP groups were 1.15,1.22 and 1.33 times higher than those in CC group,POD activity was 1.33,1.46 and 1.85 times higher than those of CC group,respectively.The content of MDA in tobacco leaves was decreased by 21.98%,23.98% and 24.84% in CP,PC and PP groups respectively.The expression levels of NTNAC1 and NAC4 in tobacco leaves were up-regulated.The expression levels of NTNAC1 in CP,PC and PP groups were 3.37,3.88 and 5.07 times higher than those in CC group,the expression levels of NAC4 gene were 3.04,3.59 and 5.56 times higher than those of CC group,respectively.This study indicates that P.indica and AMF showed significant synergistic effects,which could significantly improve the drought resistance of tobacco.

Key words: Tobacco, Piriformospora indica, Arbuscular mycorrhizal fungi, Drought resistance

孙文秀, 邵晨阳, 陈妍妍, 聂明皓, 李震, 曹毅, 刘应保. 干旱胁迫下2种内生真菌对烟草生理生化指标及NAC基因表达的影响[J]. 华北农学报, 2024, 39(1): 113-119. doi: 10.7668/hbnxb.20194526.

SUN Wenxiu, SHAO Chenyang, CHEN Yanyan, NIE Minghao, LI Zhen, CAO Yi, LIU Yingbao. Effects of Two Endophytic Fungi on Physiological and Biochemical Indexes and NAC Gene Expression in Tobacco under Drought Stress[J]. Acta Agriculturae Boreali-Sinica, 2024, 39(1): 113-119. doi: 10.7668/hbnxb.20194526.

http://www.hbnxb.net/CN/Y2024/V39/I1/113

图/表 7

表1 荧光定量PCR引物

Tab.1 Primers of fluorescent quantitative PCR

基因登录号 Gene accession number	引物名称 Primers name	引物序列(5'—3') Sequence of primers (5'—3')
NM_001325686.1	Actin For	TGGTTAAGGCTGGATTTGCT
	Actin Rev	TGCATCCTTTGACCCATAC
XM_016622840.1	NtNAC1 For	GTTTATGGGCAGAAGGAGCAG
	NtNAC1 Rev	CCTGGGTGATTTGCCTTGTTT
XM_016656095.1	NAC4 For	GGCAGGCGTTGTAGTAGGTGA
	NAC4 Rev	GATGTGGCTTGTGAATGTCTCG

图1 干旱处理5 d后烟草的生长状况

Fig.1 Growth status of tobacco after 5 days of drought treatment

表2 干旱处理7 d后烟草的生物学特性

Tab.2 Biological characteristics of tobacco plants after 7 days of drought treatment

处理 Treatment	主根长/cm Main root length	地下部分鲜质量/g Underground part fresh weight	地下部分干质量/g Underground part dry weight	地上部分鲜质量/g Aboveground part fresh weight	地上部分干质量/g Aboveground part dry weight
CC	5.18±0.52a	2.77±0.09a	0.13±0.29a	18.11±0.24b	1.24±0.16a
CP	6.73±0.09b	3.69±0.22a	0.26±0.39a	25.94±0.32b	2.51±0.27b
PC	6.51±0.17b	3.78±0.47b	0.27±0.51b	25.89±0.19c	2.53±0.44c
PP	7.69±0.03c	4.02±0.62b	0.31±0.19c	26.83±0.37c	2.80±0.26c

图2 干旱胁迫下烟草叶片中Pro含量的变化图中不同大写字母代表相同处理条件下不同时间差异显著(P<0.05),不同小写字母代表相同时间下不同处理的差异显著(P<0.05)。图3—5同。

Fig.2 Changes of Pro content in tobacco leaves under drought stress Different capital letters represent significant differences at different time under the same treatment(P<0.05),different lowercase letters represent significant differences of different treatments at the same time (P<0.05).The same as Fig.3—5.

图3 干旱胁迫下烟草叶片中SOD和POD活性的变化

Fig.3 Changes of SOD and POD activities in tobacco leaves under drought stress

图4 干旱胁迫下烟草叶片中MDA含量的变化

Fig.4 Changes of MDA content in tobacco leaves under drought stress

图5 干旱胁迫下烟草叶片中基因NTNAC1和基因NAC4相对表达量的变化

Fig.5 Changes of relative expression of NTNAC1 and NAC4 genes in tobacco leaves under drought stress

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