Disruption of OsPHD1, Encoding a UDP-Glucose Epimerase, Causes JA Accumulation and Enhanced Bacterial Blight Resistance in Rice

Lesion mimic mutants (LMMs) have been widely used in experiments in recent years for studying plant physiological mechanisms underlying programmed cell death (PCD) and defense responses. Here, we identified a lesion mimic mutant, lm212-1, which cloned the causal gene by a map-based cloning strategy, and verified this by complementation. The causal gene, OsPHD1, encodes a UDP-glucose epimerase (UGE), and the OsPHD1 was located in the chloroplast. OsPHD1 was constitutively expressed in all organs, with higher expression in leaves and other green tissues. lm212-1 exhibited decreased chlorophyll content, and the chloroplast structure was destroyed. Histochemistry results indicated that H2O2 is highly accumulated and cell death is occurred around the lesions in lm212-1. Compared to the wild type, expression levels of defense-related genes were up-regulated, and resistance to bacterial pathogens Xanthomonas oryzae pv. oryzae (Xoo) was enhanced, indicating that the defense response was activated in lm212-1, ROS production was induced by flg22, and chitin treatment also showed the same result. Jasmonic acid (JA) and methyl jasmonate (MeJA) increased, and the JA signaling pathways appeared to be disordered in lm212-1. Additionally, the overexpression lines showed the same phenotype as the wild type. Overall, our findings demonstrate that OsPHD1 is involved in the regulation of PCD and defense response in rice.

Dynamics of bacterial blight disease in resistant and susceptible rice varieties

Bacterial blight (X. oryzae pv. oryzae) is a serious disease in rice across the world. To better control the disease, it is important to understand its epidemiology and how key aspects of this (e.g. infection efficiency, and spatial spread) change according to environment (e.g. local site conditions and season), management, and in particular, variety resistance. To explore this, we analysed data on the disease progress on resistant and susceptible varieties of rice grown at four sites in the Philippines across five seasons using a combination of mechanistic modelling and statistical analysis. Disease incidence was generally lower in the resistant variety. However, we found no evidence that the primary infection efficiency was lower in resistant varieties, suggesting that differences were largely due to reduced secondary spread. Despite secondary spread being attributed to splash dispersal which is exacerbated by wind and rain, the wetter sites of Pila and Victoria in south Luzon tended to have lower infection rates than the drier sites in central Luzon. Likewise, we found spread in the dry season can be substantial and should therefore not be ignored. In fact, we found site to be a greater determinant of the number of infection attempts suggesting that other environmental and management factors had greater effect on the disease than climate. Primary infection was characterised by spatially-random observations of disease incidence. As the season progressed, we observed an emerging short-range (1.6 m–4 m) spatial structure suggesting secondary spread was predominantly short-range, particularly where the resistant variety was grown.

Identification of bacterial blight resistance genes in rice landraces from Yunnan Province, China

Bacterial blight (BB), a serious bacterial disease caused by pathogen Xanthomonas oryzae pv. oryzae (Xoo) affects rice growth and yield. Yunnan Province is regarded as a center of rice diversity in China and indeed the world, and has abundant rice landrace resources, which may offer prospective candidate donors in rice improvement and breeding. In this study, a set of 200 rice landraces were evaluated to determine their resistance to 10 pathogenic Xoo strains resistance by the leaf-clipping method. The results indicated that the tested rice landraces had different resistance levels against different Xoo strains. Multiple comparisons showed that the Xoo strain PXO99 was virulent to the tested rice landraces. Sixty-six rice landraces conferred resistance against at least one Xoo strain. These resistant rice landraces screened were then performed the presence of 14 cloned BB resistance genes by closely linked molecular markers and designed specific primers. The results showed that none of these resistant accessions contained xa13, Xa21, Xa27, and Xa45(t) homologous fragments, while 9, 24, 4, 7, 9, 15, 1, 5, 4 and 27 accessions contained Xa1, Xa2/Xa31(t), Xa14, Xa3/Xa26, Xa4, xa5, Xa7, Xa10, Xa23 and xa25 homologous fragments, respectively. Sequence analysis further revealed that nucleotide variations around functional nucleotide polymorphisms region were observed within these accessions containing the Xa1, Xa2/Xa31(t), Xa14, Xa3/Xa26, Xa4, xa5, Xa10, Xa23 and xa25 homologous fragments. These results along with phenotypic resistance spectrum supported that these accessions carried nine resistance homologous genes. Only one accession (Qishanggu_Wenshan) carried the Xa7 resistance gene. We also found that some resistant rice landraces, especially Xilandigu_Baoshan, and Laoyaling_Lincang without the above resistance genes, which mediated broad spectrum resistance to multiple Xoo strains, were identified as potential sources for breeding rice lines resistance to BB.

Avirulence gene based RFLP and rep-PCR distinguish the genetic variation of Xanthomonas oryzae pv. oryzae pathotypes in Bangladesh

Bacterial blight (BB) caused by X. oryzae pv. oryzae is one of the devastating diseases of rice mostly in Asia. Genomes of X. oryzae pv. oryzae is highly variable due to rearrangement of the large contents of transposable elements and dynamic changes of X. oryzae pv. oryzae population regulated efficiency of the control measures used for BB management of rice worldwide. In this study, genetic variation of X. oryzae pv. oryzae pathotypes of Bangladesh was studied using aviruelnce gene based RFLP and rep-PCR techniques aimed to formulate pathogen targeted effective control measures against BB of rice. Eight pathotypes of X. oryzae pv. oryzae field isolates were identified based on their reactions against 10 Near Isogenic Lines (NILs). Among eight pathotypes, pathotypes IV and V contained higher number of isolates which were 30.13% and 23.01% respectively while pathotype VIII revealed as minimum containing only 2.51% of total isolates. These eight pathotypes were studied for their genetic variation by RFLP using avrBs3 repeat domain as probe. The results conceded that Bangladeshi X. oryzae pv. oryzae strains seem carrying a minimum of two and maximum of nine avrBs3 family genes homologs. The resistance phenotype on IRBB7 and IRBB10 NILs also indicated presence of two major avrBs3 family genes viz. avrxa7 and avrXa10 in some pathotypes. Relationship of phylogenicity exhibited that X. oryzae pv. oryzae pathotypes assorted into two RFLP haplotypes as well as these haplotypes are largely distributed in Bangladesh. Phylogenetic analyses carried out by (REP, ERIC), rep-PCR and BOX depicted the presence of two main molecular haplotypes of X. oryzae pv. oryzae pathotypes. The relationship between pathotypes and molecular haplotypes of X. oryzae pv. oryzae in Bangladesh indicated that the same lineage possesses different pathotypes and different lineage possesses different pathotypes. The results indicated that eight different pathotypes might have originated from common inherited haplotypes with a wide genetic variation.

Sunflower diseases control by biological fungicides

The demand for organic food has grown rapidly over the last 25 years in many countries of the world. Organic farming standards require the application of biological products of disease control. The article presents the material on studying the effectiveness of fungicides based on microbiological antagonists against sunflower diseases. We carried out the study in 2020-2021 at V.S. Pustovoit All-Russian Research Institute of Oil Crops (VNIIMK) (Krasnodar). We established that the fungicides Geostim Fit type E, WS based on microbiological antagonists did not reduce the germination of sunflower seeds and actively suppressed seed infection. Their effectiveness against Alternaria spp. reached 50-74 %, against Rhizopus spp. – 74–100 %, against bacteria – 76–80 %. Double treatment of sunflower plants with BFTIM, WS (different strains) reduced the frequency of bacterial blight to 37 %, of dry rot – to 60 %, of Alternaria blight – to 39 %, of Phoma rot – to 41 %, and of Fusarium blight – to 75 %. The application scheme of Geostim Fit E (G), WS; BFTIM (E), WS; BFTIM (Zh), WS showed the best economic effectiveness with a seed yield of 3.56 t/ha, net income of 122 910 rubles, and profitability of 311 %.

Anatomical features of conductive tissues of soybeans that prevent the penetration of pathogenic agents of bacterial blight into seeds

The ways of penetration of pathogenic bacteria from the infected vegetative parts of plants into soybean seeds remain practically unexplored. It is widely believed that soybean seeds are infected through the vascular system from already infected areas of the vegetative parts. The aim of the present research was to study the possibility of penetration of pathogens of bacterial blight into soybean seeds through the conductive tissues of plants. The studies were carried out in 2019–2021 in V.S. Pustovoit All-Russian Research Institute of Oil Crops on plants and seeds of soybean variety Vilana. It was found that the size of stomatal slots in soybean leaves is 8–12 µm. This ensures free penetration of bacteria with a diameter of 1.3–1.7 µm into the leaf mesophyll. The pore sizes of the sieve plates of the phloem range from 0.4–0.7 to 0.8–1.6 µm, depending on the age of the plants. The largest pores of the phloem sieve plates are comparable to the diameters of pathogenic bacteria. However, a large number of transverse sieve plates located in the vessels of the phloem every 0.05–0.1 mm will filter and partially retain bacteria in each sieve tube along the path of cell sap in the phloem. Therefore, the pathogenic bacteria passing through the entire phloem from leaves infected with bacteriosis up to pods is physically unlikely. In pods, the vascular system ends in the area of attachment of the placenta to the seed hilum. In the hilum, there are no conductive tissues, and the further flow of water and nutrients into the seed is carried out diffusely through the plasmodesma of cell walls. It was found that the anatomical structure of the soybean phloem prevents the free movement of pathogenic bacteria along the conductive system directly into the inner tissues of the seeds. Therefore, the hypothesis of infection of soybean seeds with pathogens of bacterial blight through the conducting system of the plant should be considered untenable

Phenotypic Screening of F4 Breeding Lines against Bacterial Blight Disease in Rice

Biotic stresses are major threat to rice production. Among biotic stresses, bacterial leaf blight is one of the major diseases affecting rice grain production in rice growing areas. Present investigation was conducted to evaluate phenotypic effect of 50 breeding lines from a cross (Pranahitha//ISM/MTU1010) in glass house at Regional Agricultural Research Station, Jagtial by inoculating Bacterial leaf blight culture (DX-020) by leaf clipping method. Out of these 50 F4 lines evaluated, twenty nine breeding lines showed resistant reaction with disease score of 1. Eleven lines showed moderately resistant reaction with disease score 3. Twenty nine breeding lines that were resistant with disease score 1, had excellent grain yield. Hence, these lines can be advanced to further generations. Thus the present study has demonstrated that phenotypic selection is successful in the glass house and these breeding lines with higher yield levels are expected to perform better in the field trials and further in the farmers fields with the good level of bacterial blight resistance.

Susceptibility of Some Corylus avellana L. Cultivars to Xanthomonas arboricola pv. corylina

Bacterial blight of hazelnut (Corylus avellana L.) is caused by Xanthomonas arboricola pv. corylina (Xac). In the past, bacterial blight has been a key disease impacting the Oregon hazelnut industry where 99% of the United States hazelnut crop is grown. The disease is re-emerging in young orchards, as acreage of newly released hazelnut cultivars rapidly increases. This increase in hazelnut acreage is accompanied by renewed interest in developing control strategies for bacterial blight. Information on susceptibility of hazelnut cultivars to Xac is limited, partially due to lack of verified methods to quantify hazelnut cultivar response to artificial inoculation. In this research, Xac inoculation protocols were adapted to two hazelnut growing environments to evaluate cultivar susceptibility: in vitro tissue culture under sterile and controlled conditions, and in vivo potted tree conditions. Five hazelnut cultivars were evaluated using the in vitro inoculation protocol and seven hazelnut cultivars were evaluated using the in vivo inoculation protocol. Under in vitro conditions, there were severe bacterial blight symptoms on each cultivar consistent with those seen in the field, but no significant differences in the susceptibility of the newly released cultivars were observed compared to known Xac-susceptible cultivar (“Barcelona”). Under in vivo conditions, the proportion of necrotic buds were significantly higher in “Jefferson” and “Dorris” compared to all of the other tested cultivars, including “Barcelona.” The symptom progression seen in vivo mirrored the timing and symptom progression of bacterial blight reported from field observations. The in vitro conditions significantly reduced the amount of time required to measure the inoculation efficiency compared to the in vivo environment and allowed for greater replication. Further studies on the effects of Xac can use the results of these experiments to establish a dose–response model for bacterial blight, a wider range of germplasm can be tested under in vitro conditions, and management strategies that can be evaluated on large populations of new cultivars using the in vivo methods.