First Report of Root‐knot Nematodes (Meloidogyne arenaria) on Angelica dahurica in China

Angelica dahurica is an important Chinese herbal medicine plant, and its rhizome is of high medicinal value. In recent years, a severe decline in yield has been observed in Bozhou City (China's largest A. dahurica producing area), Anhui province, China. It showed symptoms of decline, stunting, yellowing and many galls in the roots, which was the characterization of infestation by root‐knot nematodes. A survey of root‐knot nematodes on its roots was conducted in this area from June to September, 2011. Based on our results, the nematode species on A. dahurica was identified as Meloidogyne arenaria by the morphological, biochemical and molecular methods. To our knowledge, this is the first report of M. arenaria on A. dahurica in China.     

Host‐Parasite Relationships in Root‐Knot Disease Caused by Meloidogyne inornata in Common Bean (Phaseolus vulgaris)

Bean crops have their productivity limited by biotic factors, as the phytonematodes. Several species have been reported causing damage to the crop, especially those from the Meloidogyne genus. Recently, a new species was reported parasitizing bean plants in Paraná State, Brazil, Meloidogyne inornata. The present work was aimed in order to study the pathosystem bean and M. inornata, through the evaluation of host reaction of 32 bean cultivars to the nematode, the potential damage of this pathogen to the crop, and the biology and parasitism of M. inornata on bean, under different temperatures, comparing with M. incognita. The host reaction was accessed under greenhouse conditions, with an initial population density of 2000 eggs of each nematode species per plant. Fifty days after inoculation, it was found that all tested cultivars were susceptible to M. inornata, however with varying extent. Pathogenicity of M. inornata on bean cv. Tuiuiú was also evaluated under greenhouse conditions, with the following initial population densities: 0 (check); 0.0625; 0.125; 0.25; 0.5; 1; 2; 4; 16; 32; and 64 eggs per cm³ of soil. Tolerance limit obtained to this cultivar was 9.9 nematodes per cm³. In relation to the comparative biology between M. inornata and M. incognita, under growth chamber and three different temperatures, 18, 25 and 32°C, results showed that under 18°C, both species have their life cycles retarded, while under 32°C, the cycle is accelerated.     

Isolation and Identification of Bacillus amyloliquefaciens IBFCBF‐1 with Potential for Biological Control of Phytophthora Blight and Growth Promotion of Pepper

In this study, 76 bacterial strains were isolated from the rhizosphere soil of pepper. Of these, 23 bacterial isolates capable of inhibiting Phytophthora capsici growth were selected. Among the antagonistic bacteria, one strain, IBFCBF‐1 showed the strongest antagonistic activity, and was identified as Bacillus amyloliquefaciens based on the results of 16S rRNA gene sequence analysis, physiological and biochemical testing, and morphological characteristics. When tested with a dual‐culture method and with laboratory greenhouse studies, the strain IBFCBF‐1 was found to be a potential biocontrol agent for controlling the plant pathogen, P. capsici. Moreover, it showed high efficiency and broad‐spectrum antifungal properties in vitro. Under greenhouse conditions, IBFCBF‐1 could significantly promote the growth of pepper seedlings, and was able to solubilize phosphate, and produce indole acetic acid (IAA) and ammonia. This study clearly demonstrated that IBFCBF‐1 is a potential candidate exhibiting phytophthora blight‐suppressive and plant growth‐promoting effects on pepper.     

Colonization ability as an indicator of enhanced biocontrol capacity—An example using two Bacillus amyloliquefaciens strains and Botrytis cinerea infection of tomatoes

An understanding of biocontrol activities is important when developing microorganism‐based alternatives to conventional fungicides. From our bacterial collection, we selected two strains (BBC023 and BBC047) for their outstanding antagonistic capacity against fungal phytopathogens and growth‐promoting abilities towards Arabidopsis thaliana. According to physiological and molecular characterizations, both strains were classified as Bacillus amyloliquefaciens and were tested against Botrytis cinerea in vitro and in a tomato. Both strains secrete lipopeptide‐like compounds that contribute to their in vitro antagonism. SEM‐images showed altered B. cinerea mycelial structures that were consistent with previous reports of the direct action of lipopeptides against fungal hyphae. The strains were applied to the roots (R), leaves (foliar ‐ F) or root/leaves (R/F) on tomato plants. All treatments significantly reduced the severity of B. cinerea infection (measured as a control index). However, only root applications (R and R/F) led to growth promotion in the tomato plants. We detected the production of indole acetic acid (IAA) and 2,3‐butanediol as growth promotion traits in the two strains. For both strains, the R/F treatment showed the highest control index, suggesting a synergic effect of direct antagonism against B. cinerea and resistance induction in the plant. In addition, in vitro antagonism of BBC023 and BBC047 against B. cinerea was similar; whereas in the F application, strain BBC047 significantly improved plant resistance and maintained a higher population density over time on tomato leaves, compared to BBC023. BBC047 was also able to produce a complex and robust biofilm in Msgg medium compared with that of BBC023. We linked the reduced biocontrol of BBC023 on leaves with its limited ability to generate robust biofilms and colonize the phylloplane. At last, we highlight the potential of the native Bacillus strains as promising alternatives for the development of bioproducts for sustainable agriculture.     

Genetic Diversity of the root‐knot nematode Meloidogyne enterolobii in Mulberry Based on the Mitochondrial COI Gene

This study explores the genetic diversity and structure of Meloidogyne enterolobii in mulberry in China. The COI mitochondrial gene (mtCOI) in M.enterolobii populations in Guangdong, Guangxi, and Hunan Provinces was PCR‐amplified, sequenced, and analyzed for genetic diversity. The total number of variations, haplotypes (Hap), the average number of nucleotide differences (k), haplotype diversity (H), and nucleotide diversity (π) of mtCOI were 25, 11, 4.248, 0.900, and 0.00596, respectively. Insignificant differences in Fst value (0.0169) and a high level of gene flow (7.02) were detected among the 19‐mulberry root‐knot nematode populations, and high genetic variation within each population and a small genetic distance among populations were observed. Both phylogenetic analyses and network mapping of the 11 haplotypes revealed a dispersed distribution pattern of 19 mulberry root‐knot nematode populations and an absence of branches strictly corresponding to the 19 range sampling sites. The neutrality test and mismatch analysis indicated that mulberry root‐knot nematode populations experienced a population expansion in the past. The analysis of molecular variance (AMOVA) revealed that the genetic differentiation of M. enterolobii was mainly contributed by the variation within each group. No significant correlation was found between the genetic distance and geographical distance of M. enterolobii populations. The findings of this study provide a profound understanding of the M. enterolobii population and will inform the development of strategies to combat and manage root‐knot nematodes in mulberry.     

The Role of Microbes Associated with Chicken Litter in the Suppression of Meloidogyne arenaria

The role of microbes associated with chicken litter in the suppression of Meloidogyne arenaria in amended soil was investigated. Amended soil treatments were prepared, including combinations of sterile and nonsterile chicken litter and soil. Microbial biomass in different treatments was compared by measuring carbon dioxide evolution. There was less CO₂ evolved in sterile litter than in nonsterile litter treatments. Tomato seedlings cv. Rutgers were transplanted into soil mixtures and inoculated with 2,000 M. arenaria eggs. After 10 days, fewer second-stage juveniles (J2) had penetrated the roots in soils amended with nonsterile litter than sterile litter. The effects of sterile and nonsterile litter-amended soil solutions on M. arenaria eggs and J2 were observed over a period of 6 days. A lower percentage of eggs remained apparently healthy in nonsterile than in sterile-amended soil solutions over 6 days. Microbial degradation of the egg shells was apparent. Fewer J2 survived in sterile- and nonsterile-amended-soil solutions as compared to water controls.     

Potential of bioinsecticidal Bacillus thuringiensis inoculum to suppress gray mold in tomato based on induced systemic resistance

The potential of the active ingredient of a commercial bioinsecticide, XenTari^® (Bacillus thuringiensis [BT] serovar aizawai strain ABTS‐1857), to suppress gray mold in tomato plants was elucidated. First, a suspension of the active ingredient of XenTari^® and a liquid culture of the bacterial strain as BT inocula were sprayed onto detached leaves or drenched into pots of tomato seedlings, and then, propagules of the gray mold fungus, Botrytis cinerea, were inoculated onto the leaves. The gray mold disease was significantly suppressed when rhizospheres were drenched with either inoculum, but not when inocula were sprayed onto detached leaves of seedlings. Both BT inocula were verified not to directly inhibit the mycelial growth of B. cinerea based on in vitro culture plate assays. Additionally, real‐time RT‐PCR analysis verified that the active ingredient increased the expression levels of defence‐related genes, such as PR‐1(P6) and P4, in the leaves of tomato seedlings. These results suggest that the active ingredient has the potential to suppress gray mold disease in tomato, not through direct antagonistic interactions with B. cinerea, but rather through systemic activation of the plant defence system by increased expression of several defence‐related genes.     

Bacillus subtilis QST 713 Confers Protection to Tomato Plants Against Pseudomonas syringae pv tomato and Induces Plant Defence‐related Genes

Bacterial speck, caused by Pseudomonas syringae pv. tomato (Pst), is an economically important disease of tomato, resulting in yield loss of marketable fruit. Management of bacterial speck is a challenge in commercial production fields due to the limited efficacy of current disease management strategies, as the pathogen acquires resistance to antibiotics and fixed copper bactericides and host resistance has not proven durable. Therefore, it is essential to develop alternative disease management strategies, like biological control. In this study, the efficacy of the commercially available biocontrol agent Bacillus subtilis QST 713 along with copper hydroxide was tested against Pst under greenhouse conditions. QST 713 reduced significantly disease severity and incidence compared to control and the copper hydroxide treatment; subsequently, the Pst population was lower in the QST 713‐treated plants compared to control. In parallel, QST 713 and copper hydroxide increased plant height compared to control and mock plants. Furthermore, the quantitative PCR analysis of PR1a, PR1b and Pin2 expression suggests a positive role for Pin2 in the plant protective activity of QST 713, as Pin2 expression was significantly higher in the QST 713‐treated plants challenged with Pst compared to the control Pst‐inoculated plants.