• Plant Biotechnology Reports
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• v.5 no.4
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• pp.301-308
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• 2011

This study tested the relative and combined efficacy of ShPx2 and ShPAL transgenes by comparing Nicotiana tabacum hybrids with enhanced levels of $_L$-phenylalanine ammonia lyase (PAL) activity and cationic peroxidase (Prx) activity with transgenic parental lines that overexpress either transgene. The PAL/Prx hybrids expressed both transgenes driven by the 35S CaMV promoter, and leaf PAL and Prx enzyme activities were similar to those of the relevant transgenic parent and seven- to tenfold higher than nontransgenic controls. Lignin levels in the PAL/Prx hybrids were higher than the PAL parent and nontransgenic controls, but not significantly higher than the Prx parent. All transgenic plants showed increased resistance to the necrotrophs Phytophthora parasitica pv. nicotianae and Cercospora nicotianae compared to nontransgenic controls, with a preponderance of smaller lesion categories produced in Prx-expressing lines. However, the PAL/Prx hybrids showed no significant increase in resistance to either pathogen relative to the Prx parental line. These data indicate that, in tobacco, the PAL and Prx transgenes do not act additively in disease resistance. Stacking with Prx did not prevent a visible growth inhibition from PAL overexpression. Practical use of ShPAL will likely require more sophisticated developmental control, and we conclude that ShPx2 is a preferred candidate for development as a resistance transgene.

• The Plant Pathology Journal
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• v.37 no.5
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• pp.428-436
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• 2021

Stem rot is a serious disease in Jerusalem artichoke (JA). To reduce the impact of this disease on yield and quality farmers often use fungicides, but this control method can be expensive and leave chemical residues. The objective of this study was to evaluate the efficacy of two biological control agents, Trichoderma harzianum T9 and Bacillus firmus BSR032 for control of Sclerotium rolfsii under field conditions. Four accessions of JA (HEL246, HEL65, JA47, and JA12) were treated or notreated with T. harzianum T9 and B. firmus BSR032 in a 4 × 2 × 2 factorial experiment in two fields (environments), one unfertilized and one fertilized. Plants were inoculated with S. rolfsii and disease was evaluated at 3-day intervals for 46 days. T. harzianum T9 and B. firmus BSR032 reduced disease incidence by 48% and 49%, respectively, whereas T. harzianum T9 + B. firmus BSR032 reduced disease incidence by 37%. The efficacy of T. harzianum T9 and B. firmus BSR032 for control of S. rolfsii was dependent on environments and genotypes. The expression of host plant resistance also depended on the environment. However, HEL246 showed consistently low disease incidence and severity index in both environments (fertilized and unfertilized). Individually, T. harzianum T9, B. firmus BSR032, or host plant resistance control stem rot caused by S. rolfsii in JA. However, no combination of these treatments provided more effective control than each alone.

• Korean Journal of Plant Tissue Culture
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• v.27 no.4
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• pp.317-323
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• 2000

Molecular genetic techniques can now be applied to the development of advanced plant genotypes, either through genetic transformation or genomic approaches which allow researchers to transfer specific traits using molecular markers. In this paper, we discuss the use of these techniques towards understanding the genetics of blackleg resistance in Brassica. In a comparative mapping study between Arabidopsis thaliana and Brassica napus, 6 R-ESTs, 7 B. napus RFLP markers and a B. napus EST were located in a collinear region of N7 (B. napus) and chromosome 1 (A. thaliana). One of the A. thaliana R-ESTs and 4 of the B. napus RFLPs co-segregated and mapped to the LmRl locus for blackleg resistance. Introgression of blackleg resistance from wild relatives is also investigated with the possibility of accelerating the introgression process via marker assisted selection.

• Plant Biotechnology Reports
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• v.2 no.1
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• pp.13-20
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• 2008

Potato (Solanum tuberosum L.), one of the most important food crops, is susceptible to a number of devastating fungal pathogens in addition to bacterial and other pathogens. Producing disease-resistant cultivars has been an effective and useful strategy to combat the attack of pathogens. Potato was transformed with Agrobacterium tumefaciens strain EHA101 harboring chitinase, (ChiC) isolated from Streptomyces griseus strain HUT 6037 and bialaphos resistance (bar) genes in a binary plasmid vector, pEKH1. Polymerase chain reaction (PCR) analysis revealed that the ChiC and bar genes are integrated into the genome of transgenic plants. Different insertion sites of the transgenes (one to six sites for ChiC and three to seven for bar) were indicated by Southern blot analysis of genomic DNA from the transgenic plants. Expression of the ChiC gene at the messenger RNA (mRNA) level was confirmed by Northern blot analysis and that of the bar gene by herbicide resistance assay. The results obviously confirmed that the ChiC and bar genes are successfully integrated and expressed into the genome, resulting in the production of bialaphos-resistant transgenic plants. Disease-resistance assay of the in vitro and greenhouse-grown transgenic plants demonstrated enhanced resistance against the fungal pathogen Alternaria solani (causal agent of early blight).

• The Plant Pathology Journal
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• v.21 no.3
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• pp.237-243
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• 2005

Resistance inductions on the leaves of cucumber plant by an arbuscular mycorrhiza Glomus intraradices were investigated. In addition, the infection structures were observed at the penetration sites on the leaves of plant inoculated with Colletotrichum orbiculare using a fluorescence microscope. The severity of anthracnose disease caused by Colletotrichum orbiculare was significantly decreased on the leaves of cucumber plant colonized with G intraradices compared with those of non-treated control plants. As a positive control, pre-treatment with DL-3-aminobutyric acid (BABA) caused a remarkable reduction of the disease severity on the pathogen-inoculated leaves. There were no significant differences in the frequency of either germination or appressorium formation of the plant pathogen between mycorrhiza colonized and non-treated plants. It was also the same on the BABA pre-treated plants. However, the frequency of callose formation was significantly high on the leaves of G intraradices colonized plants compared to those of non-treated control plants at 5 days after challenge inoculation. On the leaves of BABA treated plants callose formation was not significantly high than those of non-treated, although the disease severity was more strongly suppressed. It was suggested that the resistance induced by colonization with G. intraradices might be related to the enhancement of callose formation at the penetrate sites on the leaves invaded by the pathogen, whereas resistance by BABA did not.

• The Plant Pathology Journal
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• v.32 no.3
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• pp.228-241
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• 2016

In our previous study, we reported that a novel endophytic bacterium Bacillus oryzicola YC7007 has suppressed bacterial diseases of rice via induced systemic resistance and antibiotic production. This endophytic strain, B. oryzicola YC7007 was used as a biological control agent against bakanae disease of rice caused by Fusarium fujikuroi, and its mechanism of interaction with the pathogen and the rice was further elucidated. Root drenching with B. oryzicola YC7007 suspension reduced the disease severity of bakanae significantly when compared with the untreated controls. The treatments of B. oryzicola YC7007 suspension ($2.0{\times}10^7cfu/ml$) to the rice rhizosphere reduced bakanae severity by 46-78% in pots and nursery box tests containing autoclaved and non-autoclaved soils. Moreover, in the detached rice leaves bioassay, the development of necrotic lesion and mycelial expansion of F. fujikuroi were inhibited significantly by spraying the culture filtrate of B. oryzicola YC7007. Drenching of ethyl acetate extracts of the culture filtrate to the rhizosphere of rice seedlings also reduced the bakanae disease severity in the plant culture dish tests. With the root drenching of B. oryzicola YC7007 suspension, the accumulation of hydrogen peroxide was observed at an early stage of rice seedlings, and a hormonal defense was elicited with and without pathogen inoculation. Our results showed that the strain B. oryzicola YC7007 had a good biocontrol activity against the bakanae disease of rice by direct inhibition, and was also capable of inducing systemic resistance against the pathogen via primed induction of the jasmonic acid pathway.

• The Plant Pathology Journal
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• v.19 no.5
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• pp.253-256
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• 2003

Citrus canker disease is caused by bacteria Xanthomonas axonopodis .pv. Citri. Shiranuhi cultivar, a hybrid of Kiyomi tangor and Nakano No.3 ponkan was evaluated for resistance to citrus canker based on initiation of disease, percent area of lesion infected and growth rate of bacteria in the leaf under growth chamber condition. Significant differences between susceptible plant and resistant plants were observed in these assays. Resistant plants showed delayed disease symptoms compared to the susceptible plants after spray inoculation of the pathogen. The resistant verities, satsuma, yuzu, and Shiranuhi showed symptoms after six days where as susceptible, mexican lime showed the symptoms just after three days of inoculation. 18 days after inoculation, percent area of lesions developed on leaf and disease severity differed significantly in susceptible and resistant plants, and were ranked as follows: mexican lime > early satsuma =Shiranuhi =yuzu (P <, 0.01). However, 30 days after inoculation, percent area of lesion was further differentiated into resistant and highly resistant plants. That was ranked as follows: sweet orange> early satsuma =Shiranuhi =Kiyomi > yuzu (P < 0.01). These results indicate that host reaction to the bacterial was more distinct when the disease developed for a longer period. Growth rates of a citrus canker bacterium during 16 40 h also were distinct after infiltration into leaves of susceptible and resistant plants, and were ranked as follows: sweet orange> early satsuma =Shiranuhi =Kiyomi =yuzu (P < 0.01). Based on these results, we concluded that Shiranuhi is resistant to citrus canker as compared to Kiyomi, early satsuma, and yuzu.

• Journal of Microbiology and Biotechnology
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• v.17 no.2
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• pp.280-286
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• 2007

We evaluated a commercial biopreparation of plant growth-promoting rhizobacteria (PGPR) strains Bacillus subtilis GB03 and B. amyloliquefaciens IN937a formulated with the carrier chitosan (Bio Yield) for its capacity to elicit growth promotion and induced systemic resistance against infection by Cucumber Mosaic Virus (CMV) and Pseudomonas syringae pv. tomato DC3000 in Arabidopsis thaliana. The biopreparation promoted plant growth of Arabidopsis hormonal mutants, which included auxin, gibberellic acid, ethylene, jasmonate, salicylic acid, and brassinosteroid insensitive lines as well as each wild-type. The biopreparation protected plants against CMV based on disease severity in wild-type plants. However, virus titre was not lower in control plants and those treated with biopreparation, suggesting that the biopreparation induced tolerance rather than resistance against CMV. Interestingly, the biopreparation induced resistance against CMV in NahG plants, as evidenced by both reduced disease severity and virus titer. The biopreparation also elicited induced resistance against P. syringae pv. tomato in the wild-type but not in NahG transgenic plants, which degrade endogenous salicylic acid, indicating the involvement of salicylic acid signaling. Our results indicate that some PGPR strains can elicit plant growth promotion by mechanisms that are different from known hormonal signaling pathways. In addition, the mechanism for elicitation of induced resistance by PGPR may be pathogen-dependent. Collectively, the two-Bacilli strain mixture can be utilized as a biological inoculant for both protection of plant against bacterial and viral pathogens and enhancement of plant growth.

• The Plant Pathology Journal
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• v.33 no.4
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• pp.402-409
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• 2017

Cyclic dipeptides (CDPs) are one of the simplest compounds produced by living organisms. Plant-growth promoting rhizobacteria (PGPRs) also produce CDPs that can induce disease resistance. Bacillus vallismortis strain BS07 producing various CDPs has been evaluated as a potential biocontrol agent against multiple plant pathogens in chili pepper. However, plant signal pathway triggered by CDPs has not been fully elucidated yet. Here we introduce four CDPs, cyclo(Gly-L-Pro) previously identified from Aspergillus sp., and cyclo(L-Ala-L-Ile), cyclo(L-Ala-L-Leu), and cyclo(L-Leu-L-Pro) identified from B. vallismortis BS07, which induce disease resistance in Arabidopsis against Pseudomonas syringae infection. The CDPs do not directly inhibit fungal and oomycete growth in vitro. These CDPs require PHYTOALEXIN DEFICIENT4, SALICYLIC ACID INDUCTION DEFICIENT2, and NONEXPRESSOR OF PATHOGENESIS-RELATED PROTEINS1 important for salicylic acid-dependent defense to induce resistance. On the other hand, regulators involved in jasmonate-dependent event, such as ETHYLENE RECEPTOR1, JASMONATE RESPONSE1, and JASMONATE INSENSITIVE1, are necessary to the CDP-induced resistance. Furthermore, treatment of these CDPs primes Arabidopsis plants to rapidly express PATHOGENESIS-RELATED PROTEIN4 at early infection phase. Taken together, we propose that these CDPs from PGPR strains accelerate activation of jasmonate-related signaling pathway during infection.

• The Plant Pathology Journal
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• v.37 no.2
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• pp.194-199
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• 2021

Blackleg is a serious disease in Brassica plants, causing moderate to severe yield losses in rapeseed worldwide. Although China has not suffered from this disease yet (more aggressive Leptosphaeria maculans is not present yet), it is crucial to take provisions in breeding for disease resistance to have excellent blackleg-resistant cultivars already in the fields or in the breeding pipeline. The most efficient strategy for controlling this disease is breeding plants with identified resistance genes. We selected 135 rapeseed accessions in Sichuan, including 30 parental materials and 105 hybrids, and we determined their glucosinolate and erucic acid content and confirmed 17 double-low materials. A recently developed single-nucleotide polymorphism (SNP) marker, SNP_208, was used to genotype allelic Rlm1/rlm1 on chromosome A07, and 87 AvrLm1-resistant materials. Combined with the above-mentioned seed quality data, we identified 11 AvrLm1-resistant double-low rapeseed accessions, including nine parental materials and two hybrids. This study lays the foundation of specific R gene-oriented breeding, in the case that the aggressive Leptosphaeria maculans invades and establishes in China in the future and a robust and less labor consuming method to identify resistance in canola germplasm.