Comparative Transcriptomics and Gene Knockout Reveal Virulence Factors of Arthrinium phaeospermum in Bambusa pervariabilis × Dendrocalamopsis grandis

Arthrinium phaeospermum can cause branch wilting of Bambusa pervariabilis × Dendrocalamopsis grandis, causing great economic losses and ecological damage. A. phaeospermum was sequenced in sterile deionized water (CK), rice tissue (T1) and B. pervariabilis × D. grandis (T2) fluid by RNA-Seq, and the function of Ctf1β 1 and Ctf1β 2 was verified by gene knockout. There were 424, 471 and 396 differentially expressed genes between the T2 and CK, T2 and T1, and CK and T1 groups, respectively. Thirty DEGs had verified the change in expression by fluorescent quantitative PCR. Twenty-nine DEGs were the same as the expression level in RNA-Seq. In addition, ΔApCtf1β 1 and ΔApCtf1β 2 showed weaker virulence by gene knockout, and the complementary strains Ctf1β 1 and Ctf1β 2 showed the same virulence as the wild-type strains. Relative growth inhibition of ΔApCtf1β 1 and ΔApCtf1β was significantly decreased by 21.4% and 19.2%, respectively, by adding H2O2 compared to the estimates from the wild-type strain and decreased by 25% and 19.4%, respectively, by adding Congo red. The disease index of B. pervariabilis × D. grandis infected by two mutants was significantly lower than that of wild type. This suggested that Ctf1β genes are required for the stress response and virulence of A. phaeospermum.

Determination of Suitable Extractant for Estimating Plant Available Arsenic in Relation to Soil Properties and Predictability by Solubility-FIAM

Extractant for estimating plant available arsenic (As) in soil has not been universally established. Moreover, to assess and monitor the complex chemical behaviour of arsenic (As) in soil and subsequently its transfer in crops, a suitable extraction protocol considering the soil properties in relation to crop uptake is required. For this purpose, a pot experiment was conducted to evaluate the suitability of the extractants for determination of extractable As in soil and risk assessment by solubility-free ion activity model (FIAM) with rice (variety: Sushk Samrat) as the test crop. Soil in bulk was collected from six locations of Indo-Gangetic Plain of Bihar, India, varying in physicochemical properties to conduct the pot experiment using five doses of As (0, 10, 20, 40 and 80 mg kg−1). Six extractants namely 0.2 (M) NH4-oxalate, 0.05 (N) HCl + 0.025 (N) H2SO4, 0.5 (M) KH2PO4, 0.5 (N) NH4F, 0.5 (M) NaHCO3 and 0.5 (M) EDTA were used. The results revealed that 0.5 (M) KH2PO4 gave the best correlation with the soil properties and crop uptake and can be considered a suitable extractant of As. Regardless of the As dose and the soil type used, in rice tissue, concentration of As followed the order root > straw > leaf and grain. As high as 94% variation in As content in rice grain could be explained, when 0.5 (M) KH2PO4 extractable As is being used as input for solubility-FIAM. Extractable As cannot be determined by atomic absorption spectrophotometer (AAS) coupled with vapour generation accessory (VGA) when 0.5 (M) EDTA was used as an extractant.

Geographic Distribution, Genetic Variability and Biological Properties of Rice Orange Leaf Phytoplasma in Southeast Asia

Rice orange leaf phytoplasma (ROLP) causes clear orange to yellowish leaf discoloration and severe stunting in rice seedlings. The ecological and biological characteristics of ROLP are largely unknown because the disease has not widely caused serious problems in rice cultivated areas, thereby leading to the low accumulation of research data. However, in the past decade, the disease became a threat to rice production, particularly in South China and India; it has also been recognised in other Asian countries, such as Vietnam, Thailand and the Philippines. Here, we observed the occurrence of ROLP in paddies of the Southeast Asian counties (Cambodia, Vietnam and the Philippines) and found that the isolates in the Philippines and Vietnam were monophyletic, while those in India, Thailand and Cambodia were more diverse, suggesting their potential origins. In Cambodia, it was revealed that following polymerase chain reaction (PCR) detection, the known ROLP-insect vectors, N. virescens Distant and Recilia dorsalis Motchulsky, were ROLP-positive, indicating their roles in pathogen dispersal. Moreover, fluorescent and scanning electron microscopy revealed the intensive accumulation of the phytoplasma in phloem tissues and massive accumulation of storage starch in vascular bundle sheath and parenchyma. Altogether, this study illustrated the genetic variability of global ROLP isolates and the pathogen’s biological impact on rice tissue.

Development of rice conidiation media for Ustilaginoidea virens

Rice false smut, caused by the ascomycete Ustilaginoidea virens, is a serious disease of rice worldwide. Conidia are very important infectious propagules of U. virens, but the ability of pathogenic isolates to produce conidia frequently decreases in culture, which influences pathogenicity testing. Here, we developed tissue media with rice leaves or panicles that stimulate conidiation of U. virens. Generally, rice leaf media more effectively increased conidiation than panicle media, and certain non-filtered tissue media were better than their filtered counterparts. Among the tested media, the Indica rice leaf medium with 0.06 g/ml of Wanxian 98 leaf was most efficient for inducing conidiation, and it was also usable for conidiation-defective isolates. Although the conidia induced in rice tissue media were smaller, they were able to germinate on potato sucrose agar medium and infect rice normally. This method provides a foundation for the production of conidia in U. virens that will be widely applied in the pathogenicity testing as well as in genetic analyses for false smut resistance in rice cultivars.

Construction of a Quantitative Acetylomic Tissue Atlas in Rice (Oryza sativa L.)

PKA (protein lysine acetylation) is a key post-translational modification involved in the regulation of various biological processes in rice. So far, rice acetylome data is very limited due to the highly-dynamic pattern of protein expression and PKA modification. In this study, we performed a comprehensive quantitative acetylome profile on four typical rice tissues, i.e., the callus, root, leaf, and panicle, by using a mass spectrometry (MS)-based, label-free approach. The identification of 1536 acetylsites on 1454 acetylpeptides from 890 acetylproteins represented one of the largest acetylome datasets on rice. A total of 1445 peptides on 887 proteins were differentially acetylated, and are extensively involved in protein translation, chloroplast development, and photosynthesis, flowering and pollen fertility, and root meristem activity, indicating the important roles of PKA in rice tissue development and functions. The current study provides an overall view of the acetylation events in rice tissues, as well as clues to reveal the function of PKA proteins in physiologically-relevant tissues.