The Occurrence of Strawberry Virus 1 Infecting Strawberry in Shandong Province, China

Abstract

Strawberry (Fragaria × ananassa Duch.) is one of the most important horticultural plants worldwide with high economic and nutritional value. Strawberry virus 1 (StrV-1), closely related to the genus Cytorhabdovirus, was first reported from F. ananassa (cv. Čačanská raná) and F. vesca (cv. Rujana) in Czech Republic (Fránová et al., 2019). To identify viruses in four strawberry cultivars (cv. Honeoye, Mibao, 8128 and All Star), leaves showing virus-like symptoms (foliar chlorotic spots or streaks) were collected from Yantai in Shandong province, China in August 2020. Symptomatic leaves from two plants of each cultivar (8 samples) were pooled for high-throughput sequencing (HTS). Total RNA was extracted from the composite sample and used for constructing a cDNA library after ribosomal RNA (rRNA)-depletion. Sequencing was carried out on Illumina Hiseq 4000 (Novogene, China). Raw reads were filtered, trimmed and de novo assembled as described previously (Grabherr et al., 2013; Zhou et al. 2020). The resulting contigs were screened by BLASTn and BLASTx against GenBank database. Subsequent analyses indicated the presence of strawberry vein banding virus, strawberry pallidosis associated virus and strawberry mottle virus in the analyzed sample, which had been reported previously in strawberry (Martin and Tzanetakis, 2013; Shi et al., 2018; Bhagwat et al., 2016). Besides, five contigs ranging from 266 to 6,057 nucleotides (nt) were obtained. They shared 87 to 91% nt sequence identity with StrV-1 isolate B (GenBank accession no. MK211271). To confirm StrV-1 infection in the strawberry plants, total RNA was isolated from eight samples (used for HTS) using RNAprep Pure Plant Plus Kit (Tiangen, China). Reverse transcription polymerase chain reaction (RT-PCR) was conducted with two pairs of specific primers StrVp1 (Forward: 5ʹ-CATTACTGAAGCATTCCGTG-3′/Reverse: 5ʹ-AGATATCACGCACAGTGAC-3ʹ), and StrVp2 (Forward: 5ʹ-TTGCGCGAAGCGGATGTCCG-3′/Reverse: 5ʹ-GGCTGCCAGAGCGTTGGATG-3ʹ), targeting nt positions 70-1,231 and 7,825-9,348 of StrV-1 isolate B, respectively. Fragments with expected sizes of 1,162 bp and 1,524 bp were amplified from two samples of cv. All Star. The amplicons were cloned, sequenced, and deposited into GenBank database under accession no. MW419123, MW419124, MW645247, and MW645248. The StrV-1 sequences from All Star shared 91% to 92% nt identity with the corresponding sequences of StrV-1 isolate B. Crude sap was prepared by homogenizing the StrV-1 infected strawberry leaf tissues in 0.02 mol/L sodium phosphate buffer with 0.45% (w/v) sodium diethyldithiocarbamate thihydrate, then gently rubbed onto five healthy N. benthamiana plants. Neither the inoculated leaves nor the systemically infected leaves showed obvious symptoms seven days post inoculation. However, StrV-1 was detected by RT-PCR in all five N. benthamiana plants using RNA templates prepared from non-inoculated leaves. In addition, a survey of strawberry greenhouses was conducted in August 2020 and approximately 10% of plants in one 667 m2 greenhouse in Yantai had StrV-1 like symptoms. To the best of our knowledge, this is the first report of StrV-1 infecting strawberry in China. Our findings expand the geographic range of StrV-1 and indicate it could be a potential virus threat to strawberry production in China.

Construction of Infectious cDNA Clone of Brassica Yellows Virus Isolated from Strawberry and Establishment of TaqMan RT-qPCR

“…Strawberry viruses are causing increasingly significant harm to China’s strawberry industry as strawberry planting areas expand. Many viruses have been reported to infect strawberry plants as a result of the rapid development of HTS technology, including SPV1, CMV, TSV, and StrV-1 [ 2 , 3 , 4 , 5 ]. Previously, we discovered that single or mixed virus infection is common in strawberries [ 20 ].…”

“…However, strawberry yield and quality have been impacted by the occurrence and severity of viral diseases [ 1 ]. With the rapid development of high-throughput sequencing (HTS) technology in recent years, many viruses, including strawberry polerovirus 1 (SPV1), cucumber mosaic virus (CMV), tobacco streak virus (TSV), strawberry-associated virus 1 (SaV1), strawberry virus 1 (StrV-1), and others, have been reported to naturally infect strawberry plants [ 2 , 3 , 4 , 5 , 6 ]. Currently, approximately 30 viruses and 1 viroid have been reported to infect strawberry [ 7 ].…”

Effect of strawberry vein banding virus and strawberry mottle virus co-infection on the growth and development of strawberry

“…In China, strawberry vein banding virus (SVBV), strawberry mottle virus (SMoV), strawberry mild yellow edge virus (SMYEV), strawberry crinkle virus (SCV), strawberry necrotic shock virus (SNSV), cucumber mosaic virus (CMV), strawberry crinivirus 3 (SCrV-3), strawberry crinivirus 4 (SCrV-4) and strawberry-associated virus 1 (SaV1) have been reported to infect strawberry. (Li and Yang, 2011;Chen et al, 2014;Li et al, 2014;Chen et al, 2018;Ding et al, 2019;He et al, 2021). Only SVBV and SMoV are considered here because they are the common viruses that infect strawberry in China.…”

Symptomless infection by strawberry virus 1 (StrV‐1) leads to losses in strawberry yields

“…The virus was detected in 31% of strawberry field samples in the Czech Republic (Fránová et al, 2019). He et al (2021) found an infection rate of about 10% in samples from China, Shandong province, indicating that it may pose a potential threat to global strawberry production; however, information about its harmfulness is lacking. Therefore, we evaluated the effects of a sole StrV‐1 infection on strawberry yields and growth over a 3‐year‐long experiment using two well‐known cultivars, Darselect and Karmen.…”