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Culture of shoot tips from adventitious shoots can eradicate Apple stem pitting virus but fails in Apple stem grooving virus

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Abstract

This study attempted to eradicate Apple stem pitting virus (ASPV) and Apple stem grooving virus (ASGV) from ‘Gala’ apple using shoot tips from of adventitious shoots derived from leaf segments. Leaf segments were excised from in vitro 4-week-old stock shoots and cultured on shoot regeneration medium containing 9.1 µM thidiazuron and 0.25 µM indole-3-butyric acid (IBA) to induce adventitious shoot formation. Shoot tips of different size and different developmental stage were excised from adventitious shoots and cultured on shoot tip culture medium containing 1.1 µM 6-benzyladenine and 0.05 µM IBA. Results showed size and developmental stage of shoot tips excised from adventitious shoots did not influence survival rate, but significantly affected shoot regrowth rate and ASPV-free frequency. Shoot regrowth rates increased from 10 to 15 % in 0.3 mm shoot tips containing two leaf primordia (LP) excised after 2–3 weeks of shoot regeneration, to 53–55 % in those containing three LP excised after 3–4 weeks. The highest shoot regrowth rate (82 %) was obtained in shoot tips of 0.4 mm shoot tips containing four LP excised after 4 weeks. ASPV-free frequencies (95–100 %) were high in 0.2–0.4 mm shoot tips containing two to three LP excised after 2–4 weeks, but low (20 %) in 0.4 mm shoot tips containing four LP excised after 4 weeks. None of the shoots regenerated from the shoot tips were ASGV-free, regardless of the size and developmental stage at which shoot tips were excised. Histological studies and virus localization provided explanations to the varying frequencies of the virus eradication using different size of shoot tips that were excised from adventitious shoots at different developmental stages. The protocol described here was characteristic of high frequency ASPV eradication and high efficiency of production of shoot tips that were used for virus eradication, and thus may have potential applications to virus eradication in other plant species.

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Abbreviations

AD:

Apical dome

ApMV:

Apple mosaic virus

ASGV:

Apple stem grooving virus

ASPV:

Apple stem pitting virus

BA:

6-Benzyladenine

CMV:

Cucumber mosaic virus

GCLV:

Garlic common latent virus

GLV:

Garlic latent virus

IBA:

Indole-3-butyric acid

ISSR:

Inter-simple sequence repeat

LP:

Leaf primordia

LSV:

Lily symptomless virus

LYSV:

Leek yellow stripe virus

MD:

Meristematic dome

MS:

Murashige and Skoog medium

NAA:

Naphthalene acetic acid

OYDV:

Onion yellow dwarf virus

RAPD:

Random amplified polymorphic DNA

RT-PCR:

Reverse transcription-polymerase chain reaction

SLV:

Shallot latent virus

SRM:

Shoot regeneration medium

STCM:

Shoot tip culture medium

TDZ:

Thidiazuron

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Acknowledgments

The authors acknowledge financial support from the fund provided Department of Science and Technology of Shaanxi Province (2014KTCL02-05).

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Authors and Affiliations

  1. State Key Laboratory of Crop Stress Biology for Arid Areas, College of Horticulture, Northwest A&F University, Yangling, 712100, Shaanxi, People’s Republic of China

    Min-Rui Wang, Bai-Quan Li, Chao-Hong Feng & Qiao-Chun Wang

  2. Institute of Plant Protection, Henan Academy of Agricultural Sciences, Zhengzhou, 450002, People’s Republic of China

    Chao-Hong Feng

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  1. Min-Rui Wang
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  2. Bai-Quan Li
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  4. Qiao-Chun Wang
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Correspondence to Qiao-Chun Wang.

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Wang, MR., Li, BQ., Feng, CH. et al. Culture of shoot tips from adventitious shoots can eradicate Apple stem pitting virus but fails in Apple stem grooving virus . Plant Cell Tiss Organ Cult 125, 283–291 (2016). https://doi.org/10.1007/s11240-016-0948-y

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