First Report of Soybean Seedling Disease Caused by Rhizoctonia solani AG-7 in Taiwan

Abstract

From August to November 2020, reduced emergence and damping-off of soybean seedlings were observed in two fields (Benzhou and Wandan) in Taiwan. Disease incidence was approximately 40% in Benzhou by field scouting. The roots of damping-off seedlings were brown. Affected seedlings could be easily pulled out from the soil and the lesions on the roots/stem were generally dry and sunken. These symptoms suggested the possibility of Rhizoctonia infection. Soil surrounding symptomatic seedlings were collected to bait the potential pathogen and symptomatic plants were used for pathogen isolation. The diseased tissues were washed with tap water and surface-disinfected with 1% bleach before placing on the Dexon selection medium at 26°C for 2 days (Ko and Hora 1971). Hyphae were transferred to potato dextrose agar (PDA), and a brown colony with brown and irregular-shaped sclerotia grew from 90 out of 99 isolates. The hyphae exhibited typical characteristics of Rhizoctonia solani, including a constriction and a septum near the end of branching hyphae (Ajayi-Oyetunde and Bradely, 2018). Two isolates from Benzhou and two isolates from Wandan were tested for their pathogenicity, and eight surface-disinfected seeds were distributed evenly on the water agar plates covered by 2-day-old mycelia at 25°C in dark for 7 days. All isolates caused cotyledon rot and reduced germination. To verify their pathogenicity in pots, double-sterilized sorghum seeds were inoculated with two strains and incubated at 25°C for 2 weeks to be used as fungal inoculum (Ajayi-Oyetunde and Bradely, 2017). A layer of 15 ml of fungal inoculum was placed 5 cm beneath the soil surface in pots. Four soybean seeds were planted approximately 3 cm above the inoculum in each pot. After two weeks, reddish lesions on the hypocotyls or taproots of all seedlings in the inoculated pots were observed, while seedlings in the control pots inoculated with sterile sorghum seeds remained healthy. The pathogen was re-isolated from lesions and had identical morphology to the original isolates. To characterize the fungal identity, the internal transcribed spacer (ITS) was sequenced using the primers ITS1/ITS4 (Sharon et al., 2006). Using BLASTN in the NCBI database, the sequence (GenBank no. MW410857 and MW410858) showed 100% (639/639 bp) similarity to KF907734 and 99.83% (635/636 bp) similarity to AF354099, both belong to R. solani anastomosis group 7 (AG-7) (Hua et al. 2014; Gonzalez et al. 2001). Phylogenetic analysis comparing sequences with different AGs (Ajayi-Oyetunde and Bradely, 2017) grouped our isolates within the AG-7 clade with a 100% bootstrap confidence. In the anastomosis test, an incompatible zonation and unequal mycelial growth rates were observed when AG-7 isolates were paired with an AG-1 IA isolate. On the other hand, the compatible tuft reaction was observed when two AG-7 isolates were paired, and the compatible merge reaction was observed in the self-pairing tests (Macnish et al. 1997). Accordingly, the molecular and morphological characterizations confirmed the causal pathogen as R. solani AG-7. R. solani AG-7 was first reported on radishes in Japan (Homma et al., 1983), first found on carnation in Taiwan (Lo et al., 1990), and in field soils of various crops but not soybean (Chuang, 1997). It was suggested that Rhizoctonia diseases of soybean may be present in Taiwan, but molecular confirmation was lacking (Anonymus, 1979). As R. solani AG-7 causes diseases of soybean in the US and Japan (Baird et al., 1996), the importance of AG-7 as an endemic pathogen of soybean in Taiwan should be recognized and its prevalence determined as a first step to managing this disease.

Population Genetics and Anastomosis Group’s Geographical Distribution of Rhizoctonia solani Associated with Soybean

“…Besides, AG-2 and AG-5 have been reported to cause hypocotyl rot of soybean in Japan [ 44 ]. AG-7 is responsible for the damping-off of soybean seedlings in Taiwan [ 42 ]. In recent years, the frequency of legumes in crop rotations has increased, and also the intensive cultivation of soybean might be another reason for increasing the frequency of R. solani AG [ 4 , 5 , 21 , 42 , 61 , 62 , 63 ].…”

“…A sequence of Atheliarolfsii FSR-052 (GenBank Accession No. AY684917) (anamorph, Sclerotiumrolfsii ) was used as the outgroup [ 38 , 39 , 41 , 42 , 43 , 44 , 45 , 46 , 47 , 48 ].…”

“…Information on the genetic diversity and distribution of R. solani AG associated with soybean of a particular country is available [ 1 , 42 , 43 , 44 ]. However, there was no attempt to investigate the global genetic diversity and distribution of R. solani AG associated with soybean.…”

Heritability and gene functions associated with sclerotia formation of Rhizoctonia solani AG-7 using whole genome sequencing and genome-wide association study

“…solani AG-7 is one of the destructive AGs that can cause diseases on cotton, potato, soybean and watermelon [2,[43][44][45][46][47][48]. It has also become one of the emerging diseases for soybean production worldwide [2,10,49]. However, among many R. solani AGs that have been genome-sequenced [50][51][52][53][54][55], AG-7 is currently an orphan and genomic information is lacking.…”

“…On the other hand, AG-3 causes stem canker and dark scurf on potato (Solanum tuberosum) [5], and AG-8 causes root rot on wheat (Triticum aestivum) [6]. Specifically for soybean (Glycine max), AG-1 IA, IB and IC are the only known groups to cause web and aerial blight on leaves and leaf nodes [7,8], and another six AGs, AG-2, AG-3, AG-4, AG-5, AG-7 and AG-11, have been reported to infect below-ground parts and cause seedling diseases such as seed rot, root rot, hypocotyl lesions and pre-/post-emergence damping off [2,[9][10][11][12]. With the genetic diversity of R. solani AGs and difference in their pathogenicity preferences, management of Rhizoctonia diseases is continuously a challenge for different crops and various strategies have been proposed to provide an integrated management.…”