Molecular Detection and Sequence Analysis of ORSV and CymMV in Cultivated Cymbidium goeringii in Dali,Yunnan

doi:10.16420/j.issn.0513-353x.2020-1053

Acta Horticulturae Sinica ›› 2022, Vol. 49 ›› Issue (3): 655-662.doi: 10.16420/j.issn.0513-353x.2020-1053

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Molecular Detection and Sequence Analysis of ORSV and CymMV in Cultivated Cymbidium goeringii in Dali,Yunnan

CHEN Jianxin¹, WEI Yuqian¹, TANG Jie¹, ZHU Yongjin¹, MA Huancheng², WU Jianrong¹^,²^,^**()

¹Key Laboratory of Forest Disaster Warning and Control in Universities of Yunnan Province,College of Biodiversity Conservation,Southwest Forestry University,Kunming 650224,China
²Key Laboratory of Biodiversity Conservation in Southwest China,State Forestry Administration,College of Biodiversity Forestry,Southwest Forestry University,Kunming 650224,China

Received:2021-07-07 Revised:2021-11-25 Online:2022-03-25 Published:2022-03-25
Contact: WU Jianrong E-mail:1176279044@qq.com

Abstract

Abstract:

The survey on virus diseases was carried out and the samples of infected seedlings were collected in Dali,Yunnan,China during 2018—2019. The Reverse Transcription-Polymerase Chain Reaction （RT-PCR）was used to detect the virus. The results showed that total 70 sample were infected by ORSV （odontoglossum ringspot virus）. However,CymMV（cymbidium mosaic virus）was not detected,indicating the major potential infection thread may be from ORSV in Dali. Phylogenetic analysis based on Coat Protein gene（cp）showed that all ORSV clustered into a single clade. The results of protein analysis showed that ORSV coat protein belonged to TMV coat protein family,and the theoretical isoelectric point was 5.01,which was a hydrophilic stable protein. Thus,we confirmed that ORSV had infected in cultivated C. goeringii in Dali. The follow up study should be carried out soon on early diagnosing and screening of anti-virus medicine for orchid viral disease.

Key words: Cymbidium goeringii, CymMV, ORSV, molecular detection, cp gene, bioinformatics

CLC Number:

S682.31

CHEN Jianxin, WEI Yuqian, TANG Jie, ZHU Yongjin, MA Huancheng, WU Jianrong. Molecular Detection and Sequence Analysis of ORSV and CymMV in Cultivated Cymbidium goeringii in Dali,Yunnan[J]. Acta Horticulturae Sinica, 2022, 49(3): 655-662.

Figures/Tables 5

Table 1 Primers for RT-PCR to detect cp gene of CymMV and ORSV

引物 Primer	序列（5′-3′） Sequence	预期片段/bp Expected size	参考文献 Reference
Cy-CPF1/Cy-CPR1	ATGGGAGAGYCCACTCCARCYCCAGC/ TTCAGTAGGGGGTGCAGGCA	669	Lee and Chang,2006
OR-CPF1/OR-CPR1	ATGTCTTACACTATTACAGACCCG/ GGAAGAGGTCCAAGTAAGTCC	474	Lee and Chang,2006

Fig. 1 Symptoms of orchids infected by virus a,b：Mosaic and chlorosis;c,d：Chlorosis and necrosis;e：Streak;f：Necrosis and ringspot;g,h：Symptoms of intact plant.

Fig. 2 The RT-PCR detection of CymMV and ORSV with specific primers of cp gene M：Marker molecular weight 100-2 000 bp DNA ladder;N：Plant of symptom-free;P：Positive control;01-03：Samples of Cymbidium.

Fig. 3 Phylogenetic trees of ORSV isolates from Dali and other isolates by Neighbor-joining method a：Based on cp gene nucleotide sequence;b：Based on amino acid sequence encoded by cp gene.

Fig. 4 Analysis of protein hydrophilicity and hydrophobicity Longitudinal axis > 0 represents hydrophobic region and < 0 represents hydrophilic region.

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Molecular Detection and Sequence Analysis of ORSV and CymMV in Cultivated Cymbidium goeringii in Dali,Yunnan

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