Molecular Detection of Candidatus Liberibacter Asiaticus, the Causal Organism of Huanglongbing (Citrus Greening) in Faisalabad, Pakistan for Huanglongbing Management

Yaqub, Muhammad S.

doi:10.21162/pakjas/17.4455

Cited by 11 publications

(10 citation statements)

References 15 publications

(24 reference statements)

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“…DNA of citrus cultivars was amplified by two primer pairs, which amplified the PCR products at the desired size of ~1160bp and ~703bp, respectively. PCR product size has been confirmatory through previous literature that also reported similar band sizes using their respective primer pairs (Chohan et al, 2007;Zafarullah et al, 2016;Yaqub et al, 2017). The primer used in these studies is based on the amplification of bacterial 16s rDNA and β operon regions.…”

Section: Molecular Detectionsupporting

confidence: 76%

“…Recently, established and advanced techniques like molecular, biological, and serological assays for pathogen detection have been reviewed (Iftikhar et al, 2016;Ding et al, 2020). PCR and qPCR have been engaged for the detection of two markers, one for the 16s rDNA (OI1 and OI2C) and the other for Las (Boperon) A2/J5 (Razi et al, 2014;Yaqub et al, 2017). Correct and quick detection of a pathogen provides a base to formulate the management strategies for HLB.…”

Section: Kinnowmentioning

confidence: 99%

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Incidence and Molecular Detection of Greening Disease in Two Citrus Cultivars in Sargodha, Pakistan

Sajid¹,

Ghazanfar²,

Rauf³

et al. 2021

SJA

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C itrus is infected by several different pathogens, including fungi, prokaryotes, viruses, nematodes, and virus-like pathogens (Khan et al., 2016). Among these pathogens, the citrus greening disease is one of the most devastating and widely distributed pathogens in citrus orchards of significant citrus-growing areas of the world and Pakistan (Iftikhar et al., 2016;Saifullah et al., 2015). Citrus greening (CGD) disease is now alternatively termed as Huanglongbing (HLB) and is known with different names in different countries owing to their local languages. All the citrus cultivars are affected by this disease regardless of the rootstock. This graft transmissible disease can also be transmitted through an insect vector very efficiently (Graca, 1991;Batool et al., 2007). HLB being a century-old disease, probably originated at the end of the 19 th century in China (Batool et al., 2007; Bove, 2006). The greening disease is caused by gram-negative unculturable phloem limited bacterium (Murray and Schlifer, 1994) named Candidatus liberibacter. Jagoueix et al.

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Section: Molecular Detectionsupporting

confidence: 76%

Section: Kinnowmentioning

confidence: 99%

Incidence and Molecular Detection of Greening Disease in Two Citrus Cultivars in Sargodha, Pakistan

Sajid¹,

Ghazanfar²,

Rauf³

et al. 2021

SJA

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“…from various samples of citrus. It was observed that CLas is responsible for HLB in citrus (Ruangwong and Akarapisan, 2006;Yaqub et al, 2017) and a pair (OI1/OI2C) of primers was used for the detection of "CLas" (Munir et al, 2019). The primer amplified a product of 1160 bp but no amplicon was detected for healthy samples.…”

Section: Sequencing Of the Causal Agent Of Hlbmentioning

confidence: 99%

Prevalence and Diversity of Candidatus Liberibacter Species in Main Citrus Growing Areas of Malakand in Northwest Pakistan

et al. 2019

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Malakand Division in northwest Pakistan is known for delicious sweet orange, locally called Malta (Citrus sinensis). However, both quality and quantity of the citrus produce has seen a decline over the past few decades. Among production constraints, Huanglongbing (HLB) caused by phloem-limited alpha proteobacterium Candidatus Liberibacter asiaticus (CLas) -an endosymbiont of Asian Citrus Psyllid (Diaphorina citri), is of great economic significance. Therefore, we assessed HLB incidence through symptom-based surveys in major citrus growing areas of Khyber Pakhtunkhwa (KP), Northwest Pakistan. The presence or absence of CLas in the collected citrus samples was confirmed by PCR (Polymerase Chain Reaction) using three set of primers (LSS/LAS606, OI1/OI2C “16S rDNA” and MH353/MH354 “operon “nusG-rplK”). An average disease incidence of 92% and severity of 34% were recorded from main production zones in district Swat, KP. Likewise, in district Lower Dir, HLB had disease incidence and severity of 64% and 24%, respectively. Molecular diagnostics using 16S rDNA-specific primer LSS/LAS606 and OI1/OI2C, yielded characteristic bands of 500 bp and 1160 bp, from all five pooled samples from Swat district, while only two samples of Dir tested positive for both primers. Using an alternative primer MH353/MH354 (nusG-rplK) showed positive amplification for four samples from Swat and two samples of Dir with an amplicon of 631 bp. To assess the molecular diversity of the CLas, Sanger sequencing was carried out based on16S rDNA gene. Sequence analysis of 16S rDNA gene from all samples revealed maximum similarity with C. Liberibacter asiaticus. Among studied isolates, SwKjuPk-5 (MH374503) and SwMtaPk-2 (MH374500) isolates appeared to be more divergent and grouped in separate clad as compared to remaining isolates of Swat and Dir.

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“…Similarly, citrus plantations are also facing decline attributable to many devastating diseases. Virus and virus-like diseases of citrus crop are considered a major citrus production constraint (Yaqub et al, 2017;Fateh et al, 2017;Satir et al, 2016). Reports showed that area under citrus cultivation and its production has been increased incredibly in southern Punjab in last two decades (Table 1, 2).…”

Section: Introductionmentioning

confidence: 99%

Serological and Molecular Based Detection of Graft-Transmissible Pathogens Associated With Citrus From Non-Core Areas of Pakistan

Naqvi¹

2017

PAKJAS

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To assess prevalence of virus and virus-like diseases of citrus, surveys were conducted in citrus groves of Southern Punjab during 2013-15. Based on serological and PCR detection methods, citrus orchards and nurseries were tested for prevalence of Citrus tristeza virus (CTV), Citrus psrosis virus (CPsV), Citrus variegated chlorosis (CVC) caused by Xyellela fastidiosa, huanglongbing (HLB, citrus greening) caused by 'Candidatus Liberibacter asiaticus' and citrus stubborn disease (CSD) caused by Spiroplasma citri. Citrus leaves and twigs samples of 'Kinnow' and'Feutrell's Early' mandarins (Citrus reticulate Blanco), 'Mosambi' sweet oranges (Citrus aurantium L.), 'Shamber' grapefruit (Citrus aurantiumL.) were collected from five districts of southern Punjab viz. Multan, Khanewal, Lodhran, Layyah and Bhakkar. Direct antibody sandwich enzyme linked immuno sorbent assay (DAS-ELISA) was employed to detect CTV, CPsV, CVC and CSD, where-as HLB was detected by PCR using specific primers from two regions 16S rDNA intergenic region and ribosomal protein gene of rplKAJL-rpoBCoperon (β-operon) of 'C. Librebacter asiaticus'. Sequence of 16S rDNA revealed nucleotide identity level upto 98% among different isolates. Disease incidence of CTV, CVC, HLB and CSD was 36.7, 10, 40 and 36.7%, respectively, where-as CPsV was not detected. All 'citrus species and relatives' viz., Kinnow, sweet oranges, Feutrell's early and grapefruit (shamber) were susceptible to these diseases. To our understanding this is first investigation of virus and virus-like diseases in citrus from noncore areas of Punjab, Pakistan.

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Molecular Detection of Candidatus Liberibacter Asiaticus, the Causal Organism of Huanglongbing (Citrus Greening) in Faisalabad, Pakistan for Huanglongbing Management

Cited by 11 publications

References 15 publications

Incidence and Molecular Detection of Greening Disease in Two Citrus Cultivars in Sargodha, Pakistan

Incidence and Molecular Detection of Greening Disease in Two Citrus Cultivars in Sargodha, Pakistan

Prevalence and Diversity of Candidatus Liberibacter Species in Main Citrus Growing Areas of Malakand in Northwest Pakistan

Serological and Molecular Based Detection of Graft-Transmissible Pathogens Associated With Citrus From Non-Core Areas of Pakistan

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