PMID- 33673767
OWN - NLM
STAT- Publisher
LR - 20240222
IS - 0191-2917 (Print)
IS - 0191-2917 (Linking)
DP - 2021 Mar 5
TI - First Report of 16SrII-V Phytoplasma Associated with Green Manure Soybean
(Glycine max L.) in Taiwan.
LID - 10.1094/PDIS-12-20-2714-PDN [doi]
AB - QING PI DOU, a local variety of soybean (Glycine max (L.) Merrill) with small
seed size, is primarily cultivated in the southern region of Taiwan. Due to the
advantage of high germination rate, fast growth and high nitrogen fixation
capacity, QING PI DOU has widely used as green manure in rotation with rice to
increase soil fertility in Taiwan. In the summer of 2020, phytoplasma-induced
disease symptoms were observed in QING PI DOU with 23% (18/78) disease incidence
in Yunlin County, Taiwan. These plants exhibited severe disease symptoms such as
little leaf, yellowing, phyllody, virescence, and witches' broom compared to
healthy plants. Leaf samples of the symptomatic plants were subsequently
collected and examined through transmission electron microscopy (TEM), PCR, and
western blotting analyses. The ultrathin sections of the diseased QING PI DOU
were double-stained with uranyl acetate and lead citrate. The typical
phytoplasma-like pleomorphic bodies were observed in sieve elements of leaf veins
by TEM. To investigate the association of phytoplasma with the diseased QING PI
DOU, total DNA extracted by the Plant Genomic DNA Purification Kit (DP022,
Genemark, Taiwan) was examined by nested PCR using the phytoplasma universal
primer pair P1/P7 followed by R16F2n/R16R2 (Lee et al. 1993). The 1.2 kb PCR
product specific for 16S ribosomal RNA (16S rRNA) gene was only amplified from
symptomatic plants but not from healthy plants. BLAST analysis demonstrated that
the sequence (accession no. MW393690) of amplified DNA fragment of 16S rRNA is
identical to that of GenBank accession no. NZ_AMWZ01000008 (complement [31109 to
32640]) of peanut witches' broom (PnWB) phytoplasma, a 'Candidatus phytoplasma
aurantifolia'-related strain (Firrao et al. 2004). Further analysis on the
virtual RFLP pattern of MW393690 generated by iPhyClassifier confirmed that the
phytoplasma identified in the diseased QING PI DOU can be classified into the
16SrII-V subgroup. Samples examined by nested PCR were further selected for total
cell extracts preparation and characterized by western blotting using the
polyclonal antibody raised against the immunodominant membrane protein (Imp) of
PnWB phytoplasma (Chien et al. 2020). An expected signal of 19 kDa specific for
Imp was only detected in symptomatic plants but not in healthy plants. Moreover,
the PCR products encoding SAP11 and phyllogen, the virulence factors responsible
for phytoplasma-induced witches' broom and phyllody symptoms (Namba 2019), were
also amplified from symptomatic QING PI DOU by PCR using the primer pairs
5'-ATGGCTCCCGAAAAAAATGATAAAGG-3'/5'-TTTTTTAGAATCATCAGGCTTTTTAG-3' (0.28 kb) and
5'-ATGGATCCAAAACTTCCAGAAACT-3'/5'-GTTTTTTTCATCATTTAAATCAT-3' (0.27 kb),
respectively. Further analysis by BLAST revealed that SAP11 and phyllogen
identified in symptomatic QING PI DOU are identical with those of PnWB
phytoplasma. To the best of our knowledge, this report is the first to describe
phytoplasma-associated soybean (Glycine max L.) witches' broom disease in green
manure soybean in Taiwan.
FAU - Wang, Chien-Jui
AU - Wang CJ
AD - Institute of Biochemistry, National Chung Hsing University, Taichung, Taiwan;
kk9514095@gmail.com.
FAU - Chien, Yuanyu
AU - Chien Y
AD - Institute of Biochemistry, National Chung Hsing University, Taichung, Taiwan;
papaya1032571@gmail.com.
FAU - Liao, Pei-Qing
AU - Liao PQ
AD - Institute of Biochemistry, National Chung Hsing University, Taichung, Taiwan;
penny23penny23@gmail.com.
FAU - Chiu, Yi-Ching
AU - Chiu YC
AD - Institute of Biochemistry, National Chung Hsing University, Taichung, Taiwan;
jean31019@gmail.com.
FAU - Chen, Yuh-Kun
AU - Chen YK
AD - Department of Plant Pathology, National Chung Hsing University, Taichung, Taiwan;
ykchen@nchu.edu.tw.
FAU - Yang, Jun-Yi
AU - Yang JY
AD - Institute of Biochemistry, National Chung Hsing University, Taichung, Taiwan.
AD - Advanced Plant Biotechnology Center, National Chung Hsing University, Taichung,
Taiwan; jyang@nchu.edu.tw.
LA - eng
PT - Journal Article
DEP - 20210305
PL - United States
TA - Plant Dis
JT - Plant disease
JID - 9882809
SB - IM
OTO - NOTNLM
OT - Glycine max L.
OT - iPhyClassifier
OT - Green manure
OT - Phytoplasma
OT - Soybean
EDAT- 2021/03/07 06:00
MHDA- 2021/03/07 06:00
CRDT- 2021/03/06 05:27
PHST- 2021/03/06 05:27 [entrez]
PHST- 2021/03/07 06:00 [pubmed]
PHST- 2021/03/07 06:00 [medline]
AID - 10.1094/PDIS-12-20-2714-PDN [doi]
PST - aheadofprint
SO - Plant Dis. 2021 Mar 5. doi: 10.1094/PDIS-12-20-2714-PDN.