Agronomy and Crop Science

‘Candidatus Phytoplasma pini’-related strain MDPP, the reference strain of subgroup 16SrXXI-B, is a pathogen associated with witches’ broom disease of Pinus spp. in North America. Here, we report the first draft genome sequence of ‘Ca. Phytoplasma pini’ strain MDPP, which consists of 474,136 bases, with a G + C content of 22.22%. This information will facilitate comparative genomics of gymnosperm-infecting phytoplasmas.

ScopeThis Standard describes procedures for official control with the aim of detecting, containing and eradicating those ‘Candidatus Liberibacter’ species which are causal agents of Huanglongbing (also known as citrus greening disease) and their vectors Trioza erytreae and Diaphorina citri. NPPOs may draw on this guidance when developing national contingency plans for outbreaks of ‘Candidatus Liberibacter’ species and their vectors.Approval and amendmentFirst approved in 2019‐09.

Huanglongbing (HLB, citrus greening disease) in the major citrus-producing states of the United States is associated with Candidatus Liberibacter asiaticus (CLas), which is vectored by the Asian citrus psyllid (ACP). Surveys were conducted in Texas from 2007 to 2017 to assess the prevalence and titer of CLas in ACPs and citrus trees. ACP and citrus leaf tissue samples were collected from suspect trees in residential areas and commercial groves (orchards) and assayed for CLas by quantitative PCR. CLas detection in ACPs (2011) preceded that of citrus trees (2012) by several months. Annual incidences of CLas-positive ACPs and leaf tissue followed an exponential growth pattern over the survey period, varying from 0.03 to 28.7% in ACPs and 0.6 to 36.5% in citrus trees. There was a significant and positive relationship between the monthly incidences of CLas-positive ACP and leaf tissue samples. The proportion of HLB detection sites also increased with time, reaching 26 and 40% of commercial groves and residential sites, respectively, by 2017. Seasonal variations were observed in the incidences of CLas-positive ACPs and citrus trees such that significantly more CLas-positive ACPs and trees were recorded during the fall and winter of a given year relative to the hot summer. A temporal analysis of the class distribution of cycle threshold values revealed a trend of increased bacterial accumulation in ACPs and trees over time, with the trend more pronounced for the former than the latter host type. These findings provide a comprehensive insight into the ongoing CLas/HLB epidemic in Texas, with potential lessons for California and other citrus-producing areas where the disease is not yet established.

Citrus huanglongbing (HLB), caused by phloem-limited ‘Candidatus Liberibacter’ bacteria, is a destructive disease threatening the worldwide citrus industry. The mechanisms of pathogenesis are poorly understood and no efficient strategy is available to control HLB. Here, we used a comparative genomics screen to identify candidate microbe-associated molecular patterns (MAMPs) from ‘Ca. Liberibacter’ spp. We identified the core genome from multiple ‘Ca. Liberibacter’ pathogens, and searched for core genes with signatures of positive selection. We hypothesized that genes encoding putative MAMPs would evolve to reduce recognition by the plant immune system, while retaining their essential functions. To efficiently screen candidate MAMP peptides, we established a high-throughput microtiter plate-based screening assay, particularly for citrus, that measured reactive oxygen species (ROS) production, which is a common immune response in plants. We found that two peptides could elicit ROS production in Arabidopsis and Nicotiana benthamiana. One of these peptides elicited ROS production and defense gene expression in HLB-tolerant citrus genotypes, and induced MAMP-triggered immunity against the bacterial pathogen Pseudomonas syringae. Our findings identify MAMPs that boost immunity in citrus and could help prevent or reduce HLB infection.

‘Candidatus Liberibacter solanacearum’ is a plant pathogen affecting the families Solanaceae and Apiaceae in different parts of the world. ‘Ca. L. solanacearum’ is a Gram-negative, fastidious α-proteobacterium that is vectored by different psyllid species. Plant-pathogenic bacteria are known for interfering with the host physiology or defense mechanisms, often by secreting bacterial effectors. Effector proteins are critical for virulence; therefore, the identification of effectors could help with disease management. In this study, we characterized the Sec-translocon-dependent ‘Ca. L. solanacearum’–hypothetical protein effector 1 (Lso-HPE1). We compared this protein sequence in the different ‘Ca. L. solanacearum’ haplotypes. We predicted the signal peptide and validated its function using Escherichia coli’s alkaline phosphatase fusion assay. Agrobacterium tumefaciens-mediated transient expression in Nicotiana benthamiana demonstrated that Lso-HPE1 from ‘Ca. L. solanacearum’ haplotypes A and B were able to inhibit the induction of cell death in plants. We also compared gene expression of the Lso-HPE1- transcripts in ‘Ca. L. solanacearum’ haplotypes A and B in tomato and in the vector Bactericera cockerelli. This work validates the identification of a Sec-translocon-dependent ‘Ca. L. solanacearum’ protein possibly involved in suppression of plant cell death.

Zebra chip (ZC) disease of potato (Solanum tuberosum) is associated with infection by ‘Candidatus Liberibacter solanacearum’ (Lso). Two haplotypes of Lso—A and B—occur in the United States. Lso haplotype B is more virulent than haplotype A, causing greater disease incidence in tubers, more severe symptoms, and greater loss in tuber yield. This study assessed whether tubers from infected plants generate new infected plants the following year. The effects of both Lso haplotypes A and B on tuber resprout were examined on five potato cultivars. When compared with noninfected tubers, overall plant emergence rate from Lso A- or B-infected tubers was lower, plants emerged slower, and plants generated lower daughter tuber yields in weight and quantity. Plants generally emerged poorly from Lso B-infected tubers and produced lower daughter tuber yields than Lso A-infected tubers. Regardless of Lso treatment, all daughter tubers were asymptomatic, and only 0.3% tested positive for Lso in experiments conducted over 2 years. This suggests that plants generated from Lso A- and Lso B-infected seed potatoes with severe ZC symptoms are likely not a significant source of Lso in potato fields.