Identification of Avramr1 from Phytophthora infestans using long read and cDNA pathogen-enrichment sequencing (PenSeq)

Potato late blight, caused by the oomycete pathogen Phytophthora infestans, significantly hampers potato production. Recently, a new Resistance to Phytophthora infestans (Rpi) gene, Rpi‐amr1, was cloned from a wild Solanum species, Solanum americanum. Identification of the corresponding recognized effector (Avirulence or Avr) genes from P. infestans is key to elucidating their naturally occurring sequence variation, which in turn informs the potential durability of the cognate late blight resistance. To identify the P. infestans effector recognized by Rpi‐amr1, we screened available RXLR effector libraries and used long read and cDNA pathogen‐enrichment sequencing (PenSeq) on four P. infestans isolates to explore the untested effectors. Using single‐molecule real‐time sequencing (SMRT) and cDNA PenSeq, we identified 47 highly expressed effectors from P. infestans, including PITG_07569, which triggers a highly specific cell death response when transiently coexpressed with Rpi‐amr1 in Nicotiana benthamiana, suggesting that PITG_07569 is Avramr1. Here we demonstrate that long read and cDNA PenSeq enables the identification of full‐length RXLR effector families and their expression profile. This study has revealed key insights into the evolution and polymorphism of a complex RXLR effector family that is associated with the recognition by Rpi‐amr1

A Phytophthora sojae Glycoside Hydrolase 12 Protein Is a Major Virulence Factor during Soybean Infection and Is Recognized as a PAMP

We identified a glycoside hydrolase family 12 (GH12) protein, XEG1, produced by the soybean pathogen Phytophthora sojae that exhibits xyloglucanase and β-glucanase activity. It acts as an important virulence factor during P. sojae infection but also acts as a pathogen-associated molecular pattern (PAMP) in soybean (Glycine max) and solanaceous species, where it can trigger defense responses including cell death. GH12 proteins occur widely across microbial taxa, and many of these GH12 proteins induce cell death in Nicotiana benthamiana. The PAMP activity of XEG1 is independent of its xyloglucanase activity. XEG1 can induce plant defense responses in a BAK1-dependent manner. The perception of XEG1 occurs independently of the perception of ethylene-inducing xylanase. XEG1 is strongly induced in P. sojae within 30 min of infection of soybean and then slowly declines. Both silencing and overexpression of XEG1 in P. sojae severely reduced virulence. Many P. sojae RXLR effectors could suppress defense responses induced by XEG1, including several that are expressed within 30 min of infection. Therefore, our data suggest that PsXEG1 contributes to P. sojae virulence, but soybean recognizes PsXEG1 to induce immune responses, which in turn can be suppressed by RXLR effectors. XEG1 thus represents an apoplastic effector that is recognized via the plant’s PAMP recognition machinery.This is the publisher’s final pdf. The published article is copyrighted by the American Society of Plant Biologists and can be found at: http://www.plantcell.org/content/27/7/205

A potato late blight resistance gene protects against multiple Phytophthora species by recognizing a broadly conserved RXLR-WY effector

Species of the genus Phytophthora, the plant killer, cause disease and reduce yields in many crop plants. Although many Resistance to Phytophthora infestans (Rpi) genes effective against potato late blight have been cloned, few have been cloned against other Phytophthora species. Most Rpi genes encode nucleotide-binding domain, leucine-rich repeat-containing (NLR) immune receptor proteins that recognize RXLR (Arg-X-Leu-Arg) effectors. However, whether NLR proteins can recognize RXLR effectors from multiple Phytophthora species has rarely been investigated. Here, we identified a new RXLR-WY effector AVRamr3 from P. infestans that is recognized by Rpi-amr3 from a wild Solanaceae species Solanum americanum. Rpi-amr3 associates with AVRamr3 in planta. AVRamr3 is broadly conserved in many different Phytophthora species, and the recognition of AVRamr3 homologs by Rpi-amr3 activates resistance against multiple Phytophthora pathogens, including the tobacco black shank disease and cacao black pod disease pathogens P. parasitica and P. palmivora. Rpi-amr3 is thus the first characterized resistance gene that acts against P. parasitica or P. palmivora. These findings suggest a novel path to redeploy known R genes against different important plant pathogens

A complex resistance locus in Solanum americanum recognizes a conserved Phytophthora effector

Late blight caused by Phytophthora infestans greatly constrains potato production. Many Resistance (R) genes were cloned from wild Solanum species and/or introduced into potato cultivars by breeding. However, individual R genes have been overcome by P. infestans evolution; durable resistance remains elusive. We positionally cloned a new R gene, Rpi-amr1, from Solanum americanum, that encodes an NRC helper-dependent CC-NLR protein. Rpi-amr1 confers resistance in potato to all 19 P. infestans isolates tested. Using association genomics and long-read RenSeq, we defined eight additional Rpi-amr1 alleles from different S. americanum and related species. Despite only ~90% identity between Rpi-amr1 proteins, all confer late blight resistance but differentially recognize Avramr1 orthologues and paralogues. We propose that Rpi-amr1 gene family diversity assists detection of diverse paralogues and alleles of the recognized effector, facilitating durable resistance against P. infestans

UvVelC is important for conidiation and pathogenicity in the rice false smut pathogen Ustilaginoidea virens

ABSTRACTRice false smut disease is one of the most significant rice diseases worldwide. Ustilaginoidea virens is the causative agent of this disease. Although several developmental and pathogenic genes have been identified and functionally analyzed, the pathogenic molecular mechanisms of U. virens remain elusive. The velvet family regulatory proteins are involved in fungal development, conidiation, and pathogenicity. In this study, we demonstrated the function of the VelC homolog UvVELC in U. virens. We identified the velvet family protein UvVELC and characterized its functions using a target gene deletion-strategy. Deletion of UvVELC resulted in conidiation failure and pathogenicity. The UvVELC expression levels during infection suggested that this gene might be involved in the early infection process. UvVELC is also important in resistance to abiotic stresses, the utilization efficiency of glucose, stachyose, raffinose, and other sugars, and the expression of transport-related genes. Moreover, UvVELC could physically interact with UvVEA in yeast, and UvVELC/UvVEA double-knockout mutants also failed in conidiation and pathogenicity. These results indicate that UvVELC play a critical role in the conidiation and pathogenicity in U. virens. Functional analysis indicated that UvVELC-mediated conidiation and nutrient acquisition from rice regulates the pathogenicity of U. virens. Understanding the function of the UvVELC homolog could provide a potential molecular target for controlling rice false smut disease

Molecular Detection and Analysis of Blast Resistance Genes in Rice Main Varieties in Jiangsu Province, China

Rice blast, caused by Pyricularia oryzae, is one of the most destructive rice diseases worldwide. Using resistant rice varieties is the most cost-effective way to control the disease, and it is crucial to analyze the resistance level and the resistance genes distribution of the main varieties. In this study, we collected 119 rice main varieties in Jiangsu province and evaluated the resistance to leaf and panicle blast and found that indica rice was more resistant to rice blast than japonica rice. Moreover, we detected the distribution of 14 resistance genes (R genes) in the 119 varieties. The distribution frequencies of three R genes, Pish, Pit, and Pia, were higher than 80%, and the Pigm had the lowest distribution frequency (1.68%), followed by Pi2 (15.18%) and Pi5, Piz-t (24.37%). Combined with the multiple stepwise regression and the resistance contribution rate, eight major R genes Pita, Pi5, Pi9, Pib, Pb1, Pikm, Piz-t, and Pi2 significantly affected the resistance of rice, and we also found that six gene combinations with 100% resistance contribution rate could effectively increase the resistance of rice varieties. In summary, monitoring the resistance level of rice varieties and analyzing their resistance genes were beneficial for rice resistance breeding

Dynamics of <i>Pyricularia oryzae</i> Population Race Structures from 2003 to 2017 in Jiangsu Province, China

Rice blast, caused by Pyricularia oryzae, is one of the most destructive rice diseases worldwide. It is essential to understand the population structure and race distribution of P. oryzae for the prevention and control of rice blast. This study collected 1584 isolates across Jiangsu province from 2003 to 2017 to determine race diversity, common and dominant race structures, and resistance profiles using eight parameters from the Chinese differential cultivars (CDC) entries. Race diversity analysis indicated that the race diversity of 1584 isolates in Jiangsu province increased from 2003 to 2006 and fluctuated steadily from 2007 to 2017. Common race structure analysis showed that the common race structure of the isolates fluctuated wildly on an annual basis. Moreover, the race ZG1 was the dominant race, suggesting that the most commonly grown rice varieties in Jiangsu are highly adaptive to race ZG1. In addition, due to a higher level (>85%) of resistance to the population of isolates from 2003 to 2017, Tetep may be conformed as the most promising donor of blast resistance for resistance breeding in Jiangsu province. In summary, it is beneficial to control rice blast by continuous monitoring of the population structure and distribution of P. oryzae with the monogenic germplasm entries and by maintaining a diversity of rice varieties

Dynamics of Pyricularia oryzae Population Race Structures from 2003 to 2017 in Jiangsu Province, China

Rice blast, caused by Pyricularia oryzae, is one of the most destructive rice diseases worldwide. It is essential to understand the population structure and race distribution of P. oryzae for the prevention and control of rice blast. This study collected 1584 isolates across Jiangsu province from 2003 to 2017 to determine race diversity, common and dominant race structures, and resistance profiles using eight parameters from the Chinese differential cultivars (CDC) entries. Race diversity analysis indicated that the race diversity of 1584 isolates in Jiangsu province increased from 2003 to 2006 and fluctuated steadily from 2007 to 2017. Common race structure analysis showed that the common race structure of the isolates fluctuated wildly on an annual basis. Moreover, the race ZG1 was the dominant race, suggesting that the most commonly grown rice varieties in Jiangsu are highly adaptive to race ZG1. In addition, due to a higher level (&gt;85%) of resistance to the population of isolates from 2003 to 2017, Tetep may be conformed as the most promising donor of blast resistance for resistance breeding in Jiangsu province. In summary, it is beneficial to control rice blast by continuous monitoring of the population structure and distribution of P. oryzae with the monogenic germplasm entries and by maintaining a diversity of rice varieties