浙江农业学报 ›› 2024, Vol. 36 ›› Issue (7): 1616-1625.DOI: 10.3969/j.issn.1004-1524.20231410
韩清宇1(), 程林润2, 李月红3, 仇智灵4, 后猛5, 楼兵干1,*(
)
收稿日期:
2023-12-20
出版日期:
2024-07-25
发布日期:
2024-08-05
作者简介:
韩清宇(1999—),女,浙江杭州人,硕士研究生,研究方向为植物真菌病害。E-mail: 415772856@qq.com
通讯作者:
*楼兵干,E-mail: bglou@zju.edu.cn
HAN Qingyu1(), CHENG Linrun2, LI Yuehong3, QIU Zhiling4, HOU Meng5, LOU Binggan1,*(
)
Received:
2023-12-20
Online:
2024-07-25
Published:
2024-08-05
摘要:
近年来甘薯基腐病是浙江甘薯生产上发生的重大病害,严重影响了甘薯的产量与品质。本研究旨在发掘高抗基腐病的甘薯栽培品种,并解析抗性相关的生理指标以及抗病相关基因表达特征,为甘薯品种改良提供科学依据。本文采用伤口接种法,对60份不同甘薯种质资源进行了基腐病抗性评价,并分析了抗、感病甘薯品种接种基腐病病原菌后抗氧化酶活性、丙二醛(MDA)含量以及抗病相关基因表达水平的差异。结果表明,60份参试甘薯材料中,仅3份表现为高抗甘薯基腐病,分别为徐D1-26、徐D10-139、J7-44。10份表现为抗病、18份表现为感病、29份表现为高感。甘薯对基腐病病原菌的抗性水平与甘薯抗氧化能力、膜脂过氧化水平以及茉莉酸信号途径调控有关。接种甘薯基腐病病原菌后,高抗品种的过氧化物酶(POD)、超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性明显高于高感品种,接种第120 h,高抗品种POD和SOD活性分别是高感品种的1.81倍和3.75倍,接种后48 h,高抗品种CAT活性是高感品种的2.93倍,而高抗品种MDA含量显著低于高感品种;qRT-PCR分析表明,茉莉酸信号途径调控的抗病相关基因IbWRKY1、IbBBX24和IbNAC1在高抗品种中均表达上调,接种后168 h,其表达量分别是高感品种的2.72倍、3.15倍和1.91倍。
中图分类号:
韩清宇, 程林润, 李月红, 仇智灵, 后猛, 楼兵干. 六十份甘薯种质资源对基腐病的抗性评价与抗病相关指标分析[J]. 浙江农业学报, 2024, 36(7): 1616-1625.
HAN Qingyu, CHENG Linrun, LI Yuehong, QIU Zhiling, HOU Meng, LOU Binggan. Evaluation of resistance to foot rot and analysis of disease resistance related indexes of 60 sweet potato germplasm resources[J]. Acta Agriculturae Zhejiangensis, 2024, 36(7): 1616-1625.
病级 Grade of disease | 标准 Standard |
---|---|
0 | 无病斑Without lesion |
1 | 0 cm<病斑长度≤3 cm 0 cm<Lesion length≤3 cm |
3 | 3 cm<病斑长度≤6 cm 3 cm<Lesion length≤6 cm |
5 | 病斑长度>6 cm Lesion length>6 cm |
表1 甘薯基腐病盆栽试验病情分级标准
Table 1 Grading standard of sweetpotato foot rot in pot experiments
病级 Grade of disease | 标准 Standard |
---|---|
0 | 无病斑Without lesion |
1 | 0 cm<病斑长度≤3 cm 0 cm<Lesion length≤3 cm |
3 | 3 cm<病斑长度≤6 cm 3 cm<Lesion length≤6 cm |
5 | 病斑长度>6 cm Lesion length>6 cm |
引物 Primers | 引物序列(5’-3’) Primer sequences(5’-3’) |
---|---|
β-Actin-F | AGCAGCATGAAGATTAAGGTTGTAGCAC |
β-Actin-R | TGGAAAATTAGAAGCACTTCCTGTGAAC |
IbBBX24-Q-F | GAGCCACAGCCACCGAAAC |
IbBBX24-Q-R | AGGTCATCAACGGCCCAAG |
IbWRKY1-F | GGAATTGAGATGGCTGTAGAGTTGTTG |
IbWRKY1-R | GAAAACCCACTAACTTGCGGCTGAGA |
IbNAC1-F | CGGCCGGGGATACAAATTTGTAAGCTT |
IbNAC1-R | GAATCGGAATCCCGGCGGCATCTC |
表2 抗性相关基因荧光定量引物
Table 2 Primers of resistance-related genes used in quantitative RT-PCR
引物 Primers | 引物序列(5’-3’) Primer sequences(5’-3’) |
---|---|
β-Actin-F | AGCAGCATGAAGATTAAGGTTGTAGCAC |
β-Actin-R | TGGAAAATTAGAAGCACTTCCTGTGAAC |
IbBBX24-Q-F | GAGCCACAGCCACCGAAAC |
IbBBX24-Q-R | AGGTCATCAACGGCCCAAG |
IbWRKY1-F | GGAATTGAGATGGCTGTAGAGTTGTTG |
IbWRKY1-R | GAAAACCCACTAACTTGCGGCTGAGA |
IbNAC1-F | CGGCCGGGGATACAAATTTGTAAGCTT |
IbNAC1-R | GAATCGGAATCCCGGCGGCATCTC |
来源 Resource | 名称 Name | 类型 Typle | 病情指数DI | 抗性等级RG | ||||
---|---|---|---|---|---|---|---|---|
14 d | 21 d | 28d | 42 d | 28 d | 42 d | |||
金华市农业科学研究院 | 金20-17 Jin20-17 | 淀粉型Starch type | 0 | 2 | 2 | 6 | HR | R |
Jinhua Academy of | 济薯26 Jishu26 | 鲜食型Fresh-eating type | 0 | 2 | 8 | 10 | R | R |
Agricultural Sciences | 薯绿1号Shulv 1 | 菜用型Vegetable use type | 2 | 2 | 8 | 12 | R | S |
金徐薯69 Jinxushu 69 | 鲜食型Fresh-eating type | 4 | 8 | 12 | 16 | S | S | |
金21-16新苗Jin21-16xin | 淀粉型Starch type | 0 | 10 | 14 | 22 | S | HS | |
金20-3 Jin20-3 | 淀粉型Starch type | 4 | 8 | 16 | 22 | S | HS | |
金20-6 Jin20-6 | 淀粉型Starch type | 4 | 6 | 18 | 22 | S | HS | |
金16S1-39 Jin16S1-39 | 淀粉型Starch type | 2 | 8 | 22 | 22 | HS | HS | |
金徐薯22 Jinxushu 22 | 淀粉型Starch type | 2 | 12 | 18 | 24 | S | HS | |
金18S3-18 Jin18S3-18 | 淀粉型Starch type | 4 | 10 | 14 | 26 | S | HS | |
金17A-22 Jin17A-22 | 淀粉型Starch type | 6 | 14 | 24 | 30 | HS | HS | |
金18S2-64 Jin18S2-64 | 淀粉型Starch type | 4 | 12 | 22 | 30 | HS | HS | |
金徐薯67 Jinxushu 67 | 淀粉型Starch type | 0 | 6 | 18 | 32 | S | HS | |
烟薯25 Yanshu 25 | 鲜食型Fresh-eating type | 2 | 14 | 20 | 34 | S | HS | |
金徐薯23 Jinxushu 23 | 淀粉型Starch type | 4 | 10 | 12 | 34 | S | HS | |
金21-16假植苗Jin21-16jia | 淀粉型Starch type | 8 | 16 | 26 | 40 | HS | HS | |
金薯926 Jinshu926 | 鲜食型Fresh-eating type | 6 | 14 | 34 | 42 | HS | HS | |
金18S4-2 Jin18S4-2 | 淀粉型Starch type | 8 | 10 | 20 | 44 | HS | HS | |
金18S1-14 Jin18S1-14 | 淀粉型Starch type | 8 | 18 | 34 | 50 | HS | HS | |
金17A-39 Jin17A-39 | 淀粉型Starch type | 0 | 16 | 36 | 54 | HS | HS | |
济薯21 Jishu 21 | 淀粉型Starch type | 0 | 16 | 38 | 62 | HS | HS | |
浙薯13 Zheshu 13 | 鲜食型Fresh-eating type | 18 | 28 | 46 | 70 | HS | HS | |
徐州农业科学研究所 | J7-44 | 兼用型Dual-purpose type | 0 | 0 | 0 | 0 | HR | HR |
Xuzhou Institute of | 徐D10-139 Xu D10-139 | 兼用型Dual-purpose type | 0 | 0 | 0 | 0 | HR | HR |
Agricultural Sciences | 徐D1-26 Xu D1-26 | 兼用型Dual-purpose type | 0 | 0 | 4 | 4 | HR | HR |
徐D1-248 Xu D1-248 | 兼用型Dual-purpose type | 0 | 0 | 2 | 6 | HR | R | |
徐42 Xu 42 | 淀粉型Starch type | 0 | 0 | 4 | 6 | HR | R | |
徐D6-118 Xu D6-118 | 兼用型Dual-purpose type | 0 | 0 | 4 | 8 | HR | R | |
徐D11-80 Xu D11-80 | 兼用型Dual-purpose type | 0 | 2 | 8 | 8 | R | R | |
徐E21-21 Xu E21-21 | 兼用型Dual-purpose type | 0 | 0 | 6 | 8 | R | R | |
徐F10-311 XuF10-311 | 兼用型Dual-purpose type | 0 | 2 | 6 | 8 | R | R | |
徐D1-95 Xu D1-95 | 兼用型Dual-purpose type | 2 | 6 | 8 | 8 | R | R | |
徐紫12 Xuzi12 | 高花青素High anthocyanin | 0 | 6 | 8 | 10 | R | R | |
徐D6-111 Xu D6-111 | 兼用型Dual-purpose type | 0 | 2 | 10 | 12 | R | S | |
徐41 Xu 41 | 淀粉型Starch type | 4 | 6 | 10 | 12 | R | S | |
徐F10-295 Xu F10-295 | 兼用型Dual-purpose type | 2 | 8 | 10 | 14 | R | S | |
徐D6-140 Xu D6-140 | 兼用型Dual-purpose type | 0 | 8 | 10 | 14 | R | S | |
徐D6-24 Xu D6-24 | 兼用型Dual-purpose type | 0 | 0 | 8 | 16 | R | S | |
徐紫11 Xuzi 11 | 高花青素High anthocyanin | 0 | 2 | 12 | 16 | S | S | |
徐紫7 Xuzi 7 | 高花青素High anthocyanin | 0 | 4 | 8 | 16 | R | S | |
徐D11-49 Xu D11-49 | 兼用型Dual-purpose type | 0 | 10 | 14 | 16 | S | S | |
徐D10-152 Xu D10-152 | 兼用型Dual-purpose type | 0 | 10 | 16 | 16 | S | S | |
徐D1-243 Xu D1-243 | 兼用型Dual-purpose type | 6 | 8 | 12 | 16 | S | S | |
徐紫10 Xuzi 10 | 高花青素High anthocyanin | 0 | 2 | 8 | 18 | R | S | |
J3-24 | 兼用型Hual-purpose type | 6 | 8 | 12 | 18 | S | S | |
J10-68 | 兼用型Hual-purpose type | 0 | 0 | 16 | 18 | S | S | |
徐紫8 Xuzi 8 | 高花青素High anthocyanin | 0 | 0 | 12 | 20 | S | S | |
徐37 Xu 37 | 淀粉型Starch type | 0 | 10 | 18 | 20 | S | S | |
J9-25 | 兼用型Dual-purpose type | 0 | 8 | 14 | 20 | S | S | |
D10-81 | 兼用型Dual-purpose type | 0 | 12 | 18 | 22 | S | HS | |
徐B21-38 Xu B21-38 | 兼用型Dual-purpose type | 8 | 12 | 20 | 22 | S | HS | |
K4-1 | 兼用型Dual-purpose type | 0 | 0 | 18 | 24 | S | HS | |
徐A6-30 Xu A6-30 | 兼用型Dual-purpose type | 0 | 10 | 22 | 24 | HS | HS | |
E21-46 | 兼用型Dual-purpose type | 6 | 12 | 26 | 28 | HS | HS | |
徐E21-74 Xu E21-74 | 兼用型Dual-purpose type | 8 | 14 | 22 | 28 | HS | HS | |
徐28 Xu 28 | 淀粉型Starch type | 4 | 12 | 26 | 28 | HS | HS | |
J12-33 | 兼用型Dual-purpose type | 0 | 18 | 26 | 28 | HS | HS | |
徐F6-36 Xu F6-36 | 兼用型Dual-purpose type | 8 | 18 | 30 | 34 | HS | HS | |
F10-23 | 兼用型Dual-purpose type | 10 | 24 | 36 | 38 | HS | HS | |
J10-49 | 兼用型Dual-purpose type | 10 | 28 | 38 | 42 | HS | HS |
表3 六十份甘薯种质资源基腐病抗性评价
Table 3 Resistance evaluation of 60 sweetpotato germplasm resources
来源 Resource | 名称 Name | 类型 Typle | 病情指数DI | 抗性等级RG | ||||
---|---|---|---|---|---|---|---|---|
14 d | 21 d | 28d | 42 d | 28 d | 42 d | |||
金华市农业科学研究院 | 金20-17 Jin20-17 | 淀粉型Starch type | 0 | 2 | 2 | 6 | HR | R |
Jinhua Academy of | 济薯26 Jishu26 | 鲜食型Fresh-eating type | 0 | 2 | 8 | 10 | R | R |
Agricultural Sciences | 薯绿1号Shulv 1 | 菜用型Vegetable use type | 2 | 2 | 8 | 12 | R | S |
金徐薯69 Jinxushu 69 | 鲜食型Fresh-eating type | 4 | 8 | 12 | 16 | S | S | |
金21-16新苗Jin21-16xin | 淀粉型Starch type | 0 | 10 | 14 | 22 | S | HS | |
金20-3 Jin20-3 | 淀粉型Starch type | 4 | 8 | 16 | 22 | S | HS | |
金20-6 Jin20-6 | 淀粉型Starch type | 4 | 6 | 18 | 22 | S | HS | |
金16S1-39 Jin16S1-39 | 淀粉型Starch type | 2 | 8 | 22 | 22 | HS | HS | |
金徐薯22 Jinxushu 22 | 淀粉型Starch type | 2 | 12 | 18 | 24 | S | HS | |
金18S3-18 Jin18S3-18 | 淀粉型Starch type | 4 | 10 | 14 | 26 | S | HS | |
金17A-22 Jin17A-22 | 淀粉型Starch type | 6 | 14 | 24 | 30 | HS | HS | |
金18S2-64 Jin18S2-64 | 淀粉型Starch type | 4 | 12 | 22 | 30 | HS | HS | |
金徐薯67 Jinxushu 67 | 淀粉型Starch type | 0 | 6 | 18 | 32 | S | HS | |
烟薯25 Yanshu 25 | 鲜食型Fresh-eating type | 2 | 14 | 20 | 34 | S | HS | |
金徐薯23 Jinxushu 23 | 淀粉型Starch type | 4 | 10 | 12 | 34 | S | HS | |
金21-16假植苗Jin21-16jia | 淀粉型Starch type | 8 | 16 | 26 | 40 | HS | HS | |
金薯926 Jinshu926 | 鲜食型Fresh-eating type | 6 | 14 | 34 | 42 | HS | HS | |
金18S4-2 Jin18S4-2 | 淀粉型Starch type | 8 | 10 | 20 | 44 | HS | HS | |
金18S1-14 Jin18S1-14 | 淀粉型Starch type | 8 | 18 | 34 | 50 | HS | HS | |
金17A-39 Jin17A-39 | 淀粉型Starch type | 0 | 16 | 36 | 54 | HS | HS | |
济薯21 Jishu 21 | 淀粉型Starch type | 0 | 16 | 38 | 62 | HS | HS | |
浙薯13 Zheshu 13 | 鲜食型Fresh-eating type | 18 | 28 | 46 | 70 | HS | HS | |
徐州农业科学研究所 | J7-44 | 兼用型Dual-purpose type | 0 | 0 | 0 | 0 | HR | HR |
Xuzhou Institute of | 徐D10-139 Xu D10-139 | 兼用型Dual-purpose type | 0 | 0 | 0 | 0 | HR | HR |
Agricultural Sciences | 徐D1-26 Xu D1-26 | 兼用型Dual-purpose type | 0 | 0 | 4 | 4 | HR | HR |
徐D1-248 Xu D1-248 | 兼用型Dual-purpose type | 0 | 0 | 2 | 6 | HR | R | |
徐42 Xu 42 | 淀粉型Starch type | 0 | 0 | 4 | 6 | HR | R | |
徐D6-118 Xu D6-118 | 兼用型Dual-purpose type | 0 | 0 | 4 | 8 | HR | R | |
徐D11-80 Xu D11-80 | 兼用型Dual-purpose type | 0 | 2 | 8 | 8 | R | R | |
徐E21-21 Xu E21-21 | 兼用型Dual-purpose type | 0 | 0 | 6 | 8 | R | R | |
徐F10-311 XuF10-311 | 兼用型Dual-purpose type | 0 | 2 | 6 | 8 | R | R | |
徐D1-95 Xu D1-95 | 兼用型Dual-purpose type | 2 | 6 | 8 | 8 | R | R | |
徐紫12 Xuzi12 | 高花青素High anthocyanin | 0 | 6 | 8 | 10 | R | R | |
徐D6-111 Xu D6-111 | 兼用型Dual-purpose type | 0 | 2 | 10 | 12 | R | S | |
徐41 Xu 41 | 淀粉型Starch type | 4 | 6 | 10 | 12 | R | S | |
徐F10-295 Xu F10-295 | 兼用型Dual-purpose type | 2 | 8 | 10 | 14 | R | S | |
徐D6-140 Xu D6-140 | 兼用型Dual-purpose type | 0 | 8 | 10 | 14 | R | S | |
徐D6-24 Xu D6-24 | 兼用型Dual-purpose type | 0 | 0 | 8 | 16 | R | S | |
徐紫11 Xuzi 11 | 高花青素High anthocyanin | 0 | 2 | 12 | 16 | S | S | |
徐紫7 Xuzi 7 | 高花青素High anthocyanin | 0 | 4 | 8 | 16 | R | S | |
徐D11-49 Xu D11-49 | 兼用型Dual-purpose type | 0 | 10 | 14 | 16 | S | S | |
徐D10-152 Xu D10-152 | 兼用型Dual-purpose type | 0 | 10 | 16 | 16 | S | S | |
徐D1-243 Xu D1-243 | 兼用型Dual-purpose type | 6 | 8 | 12 | 16 | S | S | |
徐紫10 Xuzi 10 | 高花青素High anthocyanin | 0 | 2 | 8 | 18 | R | S | |
J3-24 | 兼用型Hual-purpose type | 6 | 8 | 12 | 18 | S | S | |
J10-68 | 兼用型Hual-purpose type | 0 | 0 | 16 | 18 | S | S | |
徐紫8 Xuzi 8 | 高花青素High anthocyanin | 0 | 0 | 12 | 20 | S | S | |
徐37 Xu 37 | 淀粉型Starch type | 0 | 10 | 18 | 20 | S | S | |
J9-25 | 兼用型Dual-purpose type | 0 | 8 | 14 | 20 | S | S | |
D10-81 | 兼用型Dual-purpose type | 0 | 12 | 18 | 22 | S | HS | |
徐B21-38 Xu B21-38 | 兼用型Dual-purpose type | 8 | 12 | 20 | 22 | S | HS | |
K4-1 | 兼用型Dual-purpose type | 0 | 0 | 18 | 24 | S | HS | |
徐A6-30 Xu A6-30 | 兼用型Dual-purpose type | 0 | 10 | 22 | 24 | HS | HS | |
E21-46 | 兼用型Dual-purpose type | 6 | 12 | 26 | 28 | HS | HS | |
徐E21-74 Xu E21-74 | 兼用型Dual-purpose type | 8 | 14 | 22 | 28 | HS | HS | |
徐28 Xu 28 | 淀粉型Starch type | 4 | 12 | 26 | 28 | HS | HS | |
J12-33 | 兼用型Dual-purpose type | 0 | 18 | 26 | 28 | HS | HS | |
徐F6-36 Xu F6-36 | 兼用型Dual-purpose type | 8 | 18 | 30 | 34 | HS | HS | |
F10-23 | 兼用型Dual-purpose type | 10 | 24 | 36 | 38 | HS | HS | |
J10-49 | 兼用型Dual-purpose type | 10 | 28 | 38 | 42 | HS | HS |
图1 部分不同抗性甘薯种质资源接种GLTT01后发病情况 每幅小图从左到右依次为CK、接种14 d、接种28 d、接种42 d。
Fig.1 The symptoms of some different resistant sweet potato germplasm resources inoculated with GLTT01 From left to right in each subfigure were CK, 14 d, 28 d, 42 d after inoculation respectively.
图2 高抗/高感甘薯品种接种GLTT01后叶片抗氧化酶活性和丙二醛含量变化 J7-44是高抗品种,金18S1-14是高感品种。下同。
Fig.2 Changes of activities of antioxidant enzymes and malondialdehyde content in HR/HS sweet potato varieties after inoculation with GLTT01 Data was detected based on fresh weight from J7-44 (HR) and Jin18S1-14 (HS). The same as below.
图3 接种GLTT01后不同时间甘薯叶片中IbWRKY1、IbBBX24和IbNAC1的表达水平
Fig.3 The expression levels of IbWRKY1, IbBBX24 and IbNAC1 in sweet potato leaves inoculated with GLTT01
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