番木瓜PRSV和PLDMV复合侵染中呈中性互作

doi:10.16420/j.issn.0513-353x.2021-0804

摘要/Abstract

摘要：

商业化的抗番木瓜环斑病毒（papaya ringspot virus，PRSV）的转基因番木瓜品种‘华农1号’的应用成功控制了华南地区该毁灭性病毒。然而近年来在广东和海南的一些转基因或非转基因植株上发现了另一种具有严重危害的病毒——番木瓜畸形花叶病毒（papaya leaf distortion mosaic virus，PLDMV）。为了明确这两种病毒在番木瓜上的相互关系，通过人工接种分析二者在转基因和非转基因番木瓜上的侵染率、症状、细胞病理变化以及病毒积累量等特征。结果表明，‘华农1号’对PRSV有很强的抗性，但对PLDMV却高度感病，且PLDMV并不能协助PRSV克服转基因番木瓜对其的抗性;两种病毒单独侵染和复合侵染非转基因番木瓜均能引致严重的症状，二者症状无明显差异。细胞病理学分析表明，病毒侵染后叶片叶绿体萎缩且排列不紧凑，复合侵染并未对叶肉细胞造成更为严重的损害。接种PRSV的转基因番木瓜15 d后PRSV积累量逐渐减少，直至检测不到;而接种PLDMV后其积累量逐渐升高并保持在一个较高的水平。在转基因和非转基因番木瓜上，PRSV的积累量始终低于PLDMV，在非转基因番木瓜上，两种病毒长期共存，并各自维持相应的病毒含量。这些结果表明，抗PRSV的转基因番木瓜‘华农1号’不抗PLDMV;PRSV和PLDMV复合侵染没有在番木瓜植株中发生协生关系，表现为中性互作。

关键词: 番木瓜, 番木瓜环斑病毒, 番木瓜畸形花叶病毒, 复合侵染, 互作分析

Abstract:

The commercialized transgenic papaya‘Huanong 1’has been utilized to successfully control the destructive virus-papaya ringspot virus（PRSV）in South China. However，another new emerging virus，papaya leaf distortion mosaic virus（PLDMV）was found in some PRSV-resistant transgenic and other non-transgenic plants in Guangdong and Hainan Provinces through a field investigation. The survey results indicated that the two viruses had co-infections on some papaya plants. To clarify the interrelationship between the two viruses on papaya，the characteristics of both virus incidence，symptoms and cellular pathology，and virus accumulation on transgenic and non-transgenic papaya were analyzed by artificial inoculation. The results showed that the transgenic papaya‘Huanong 1’was still strong resistant to PRSV，but highly susceptible to PLDMV，and PLDMV could not help PRSV overcome its resistance to transgenic papaya. In addition，it was not observed significant symptomatic difference on the transgenic and non-transgenic papaya by the single- or co-infection of two viruses. The virions of the two viruses mostly existed in different mesophyll cells，which were not more seriously damaged with co-infection of two viruses compared with single viral infection. Moreover，the accumulation of PRSV gradually decreased until it was undetectable 15 days post inoculation on the transgenic papaya，while the accumulation of PLDMV gradually increased and remained at a higher level. On transgenic or non-transgenic papaya，the accumulation of PRSV virus was lower than that of PLDMV，but the two viruses co-existed for a long time and each maintained the corresponding viral contents on the non-transgenic papaya. These results indicate that transgenic papaya‘Huanong 1’is highly susceptible to PLDMV，and PRSV and PLDMV were shown as neutral with no synergistic infected in papaya plants.

Key words: Carica papaya, papaya ringspot virus, papaya leaf distortion mosaic virus, co-infection, interaction analysis

中图分类号:

S668.2

莫翠萍, 吴自林, 郭泳仪, 张曙光, 李鹏飞, 李华平. 番木瓜PRSV和PLDMV复合侵染中呈中性互作[J]. 园艺学报, 2022, 49(11): 2336-2346.

MO Cuiping, WU Zilin, GUO Yongyi, ZHANG Shuguang, LI Pengfei, LI Huaping. The Neutral Interaction of Co-infection Papaya Between Papaya Ringspot Virus and Papaya Leaf Distortion Mosaic Virus[J]. Acta Horticulturae Sinica, 2022, 49(11): 2336-2346.

图/表 6

表1 番木瓜病毒检测引物

Table 1 Primer for RT-PCR and RT-qPCR of papaya virus

病毒 Virus	引物 Name	引物序列 Primer sequence	大小/bp Size
PRSV	PRSV	F：CTTCAACAGGGCTTTCTTGC;R：GAATTACCAAGAACAATGTG	927
PRSV	PRSV-q1	F：GACATATCTGGTGTCTGGGTAATGATG;R：GCGGCATGTACCTCTCAGTT	128
PLDMV	PLDMV	F：TCCGCTCTTGATGCTGGCAAATCC;R：CGGTGAGGTTCCGTTGAATAC	652
PLDMV	PLDMV-q1	F：GGAAAACGCAAAGCCCACTT;R：CGGTGAGGTTCCGTTGAATAC	169
chy	chy	F：ATCTACAATCTTGCTAACCCTA;R：AGTCATCTTGAGAATAACCCAC	281
chy	Chy-q	F：CGGATCCTCCCATTTCCCTT; R：CCCACGGTGTAAAAATCAGCAG	151

图1 PRSV和PLDMV侵染番木瓜植株的部分样品RT-PCR的检测结果 M：DL2000 plus;chy：番木瓜内参基因。

Fig. 1 RT-PCR detection of PRSV and PLDMV from some infected samples of papaya plants M：DL2000 plus marker;chy：Internal reference gene of papaya.

表2 番木瓜‘华农1号’和‘巴西’接种PRSV和PLDMV后的侵染率

Table 2 Statistical table of incidence of PRSV and PLDMV on‘Huanong 1’and‘Baxi’papaya %

处理 Treatment	华农1号Huanong 1		巴西Baxi
处理 Treatment	PRSV	PLDMV	PRSV	PLDMV
Mock（健康对照Healthy control）	0 b	0 b	0 b	0 b
PRSV	8.70 ±1.30 a	0 b	94.07 ± 3.23 a	0 b
PLDMV	0 b	94.07 ± 3.23 a	0 b	100 ± 0 a
PRSV + PLDMV	3.61 ± 1.95 b	96.28 ± 3.72 a	97.22 ± 2..78 a	98.61 ± 1.39 a
PRSV→PLDMV	0 b	96.76 ± 1.67 a	90.74 ± 9.26 a	100 ± 0 a
PLDMV→PRSV	1.85 ± 1.85 b	100 ± 0 a	100 ± 0 a	100 ± 0 a

图2 番木瓜接种PRSV和PLDMV后的症状表现

Fig. 2 Symptomatic of papaya inoculated with PRSV and PLDMV under different treatments

图3 番木瓜接种PRSV和PLDMV后病毒积累量动态变化

Fig. 3 Changes of virus accumulation after inoculation with PRSV and PLDMV

图4 番木瓜PRSV和PLDMV单独及复合侵染60 d叶肉细胞的透射电镜图 A1、A2：相同生育期健康番木瓜叶肉细胞（对照）;B1、B2：PRSV单独侵染，叶绿体排列不紧凑;C1、C2：PLDMV单独侵染，叶肉细胞形状不规则，叶绿体萎缩且不紧凑，可见到典型的风轮状内含体和线状病毒粒子;D1、D2：复合侵染，叶绿体萎缩不紧凑，细胞被挤压形状不规则，有大量的风轮状内含体。M：线粒体;N：细胞核;CH：叶绿体;CW：细胞壁;ER：内质网;PW：风轮状体;箭头：线状病毒粒子。

Fig. 4 The cytopathology of mesophyll cell from papaya plants infected with PRSV or PLDMV and co-infected with both viruses at 60 dpi A：Mesophyll cells from healthy papaya at the same growth duration;B：Mesophyll cells from papaya infected with PRSV，chloroplast is arranged separately;C：Mesophyll cells from papaya infected with PLDMV，the shape of mesophyll cells is irregular，and they fill with the typical pinwheels and filamentous virus particles;D：Mesophyll cells from papaya co-infected with both viruses，chloroplast atrophy is not compact，the shape of cells are extruded irregular with a lot of pinwheels-like inclusions. M：Mitochondria;N：Nucleus;CH：Chloroplast;CW：Cell wall;ER：Endoplasmic reticulum;PW：Pinwheel;arrow：Filamentous virus particles.

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