Identification of Potential Vectors and Alternative Plant Hosts for the Phytoplasma Associated with Napier Grass Stunt Disease in Ethiopia

Napier grass ( Pennisetum purpureum ), the most important forage crop in East Africa, has recently been affected by a devastating disease named Napier Grass Stunt (NGS). A phytoplasma of group 16SrI has been associated with NGS in Kenya and Uganda, whereas in Ethiopia, group 16SrIII was previously identified in NGS affected fields. However, no insect vectors or alternative hosts have been recorded for NGS in East Africa. During 2005, surveys were conducted at NGS-affected plantations of Debre-Zeit and Zwai field stations in Addis Ababa. Leaf samples were collected from weeds located in and surrounding the NGS-affected areas. Leafhopper species were also surveyed by vacuum sampling in a search for natural phytoplasma vectors. Total DNA was extracted from plants and insects, and used as a template in nested polymerase chain reaction (nPCR) with universal 16S rRNA phytoplasma primers. Restriction fragment length polymorphism (RFLP), sequencing of PCR products and phylogenetic analysis were conducted for a finer identification and characterization of the phytoplasma associated with NGS. A 16SrIII-A phytoplasma with 100% of identity in the 16S rRNA sequence with that of the previously identified one in Napier Grass (accession no. DQ305977 ) was identified from alfalfa, Medicago sativa (accession no. DQ305982 ), Cynodon dactylon (accession no. DQ3058983 ), Exitianus sp. ( DQ305980 ) and Leptodelphax dymas collected in Debre Zeit (accession no. DQ305979 ) and Zwai (accession no. DQ305978 ). These findings suggest that M. sativa and Cy. dactylon are alternative reservoirs, and Exitianus sp. and L. dymas , potential vectors of the 16SrIII-A phytoplasma, which may have epidemiological implications in spreading NGS in Ethiopia.     

Assessment of the potential of Napier grass (Pennisetum purpureum) varieties as trap plants for management of Chilo partellus

Ten Napier grass [Pennisetum purpureum Schumach (Poaceae)] varieties, used in various parts of Kenya as animal fodder, were tested for their potential role as a trap crop in the management of the gramineous spotted stemborer, Chilo partellus (Swinhoe) (Lepidoptera: Crambidae), through habitat manipulation. Oviposition preference and larval survival and development were determined for each of these varieties under laboratory and screen‐house conditions. Two‐choice tests revealed that seven of the varieties tested were preferentially chosen by gravid female moths for oviposition over a susceptible maize cv. Inbred A. Larval survival was significantly lower on the majority of the Napier grass varieties (about 10%) than on maize cv. Inbred A (about 60%). Similarly, larval development was about 2 weeks longer on the majority of the Napier grass varieties. It was concluded that four of the varieties tested (Bana, Ex‐Machakos, Gold Coast, and Ex‐Nyanza‐2) had potential for use as trap crops in C. partellus management because they were more preferred by the borer moths for oviposition and allowed minimal survival of the larvae. This minimal larval survival is desirable in the conservation of C. partellus natural enemies when the cultivated host plant is out of season.     

Maize more attractive than Napier grass as non-host plants for Bactrocera cucurbitae and Dacus demmerezi

Some species of fruit flies (Diptera: Tephritidae) shelter and roost in non-host plants surrounding crops. The aim of this study was to compare the attractiveness of two plants (Poaceae): maize, Zea mays (Linné) and Napier grass, Pennisetum purpureum (Schumacher) for two species of Dacini damaging cucurbits in Reunion Island: Bactrocera cucurbitae (Coquillett) (Diptera: Tephritidae) and Dacus demmerezi (Bezzi) (Diptera: Tephritidae). Plants of the two plant species were established in pots and presented to adult flies in field cages. In each cage, a cohort consisting of adults of a given species, of known sex and age, was released. For each species, the experiment was replicated four times. The number of adult flies on the different plants as well as their location on the plant was recorded. Maize was more attractive to adults of B. cucurbitae and D. demmerezi than Napier grass irrespective of sex and sexual maturity. Most adults of both species were found under the leaves of maize and Napier grass. The effects of the date, time of day, age, and sex of flies on their attraction to plants are discussed. A possible application of this study would be to use maize as a border plant to apply bait for the control of fruit flies.     

Oviposition acceptance and larval development of Chilo partellus stemborers in drought‐stressed wild and cultivated grasses of East Africa

Maternal host choices during oviposition by herbivorous insects determine the fitness of their offspring and may be influenced by environmental changes that can alter host‐plant quality. This is of particular relevance to ‘push‐pull’ cropping systems where host preferences are exploited to manage insect pest populations. We tested how drought stress in maize and companion plants that are used in these systems affect oviposition preference, larval feeding, and development of the spotted stemborer, Chilo partellus Swinhoe (Lepidoptera: Crambidae). Five host species were tested (all Poaceae): maize (Zea mays L.), Napier grass (Pennisetum purpureum Schumach), signal grass [Brachiaria brizantha (A. Rich) Stapf], Brachiaria cv. ‘Mulato’, and molasses grass [Melinis minutiflora (Beauv.)]. Under drought stress, maize experienced as much oviposition as control unstressed maize in choice and no‐choice experiments. Similarly, larval leaf damage was not significantly different in drought‐stressed vs. unstressed maize. In contrast, oviposition occurred less on drought‐stressed than on unstressed Napier and signal grass. Oviposition acceptance and leaf damage remained low in both drought‐stressed and unstressed molasses grass and Mulato. Larval survival and development remained high in drought‐stressed maize, but not in Napier, signal, and molasses grass and Mulato, where survival and development were low in both drought‐stressed and unstressed plants. Our results indicate that herbivore responses to drought‐stressed plants depend on the plant species and that drought stress can change host preference and acceptance rankings. In particular, trap‐crops such as Napier grass may not divert oviposition from the main maize crop under drought stress conditions.     

Detection of phytoplasma by loop-mediated isothermal amplification of DNA (LAMP)

Napier stunt phytoplasma (16SrXI and 16SrIII) in eastern Africa is a serious threat to the expansion of Napier grass (Pennisetum purpureum) farming in the region, where it is widely cultivated as fodder in zero grazing livestock systems. The grass has high potential for bio-fuel production, and has been adopted by farmers as a countermeasure to cereal stem borer Lepidoptera, since it attracts and traps the insect. Diagnosis of stunt phytoplasma have been largely by nested polymerase chain reaction (nPCR) targeting the 16S rRNA gene. However, the method is laborious, costly and technically demanding. This investigation has developed a simpler but effective phytoplasma diagnostic tool, called; loop-mediated isothermal amplification of DNA (LAMP). The assay was tested on 8 symptomatic and 8 asymptomatic plants, while its detection limit was compared to nested PCR using samples serially diluted from 3 ng/μl to 0.38 pg/μl. Molecular typing of LAMP products was determined by BsrI restriction digestion and Southern blot analysis. The assay sensitivity, positive and negative predictive values were estimated, while the specificity was tested on 11 phytoplasma groups. LAMP was specific to 5 phytoplasma groups: 16SrVI, X, XI and XVI. BsrI restriction digestion produced two predicted fragments, and there was specific binding of probe DNA to the LAMP amplicons in Southern blot analysis. The assay sensitivity was 100%, while the positive and negative predictive values were 63 and 100% respectively. LAMP was 20-fold more sensitive than nested PCR. This study validates LAMP for routine diagnosis of Napier stunt and other closely related phytoplasmas.     

Host plant selection behaviour of Chilo partellus and its implication for effectiveness of a trap crop

Female lepidopterans can display a hierarchy of preference among potential host species, a trait thought to arise from the balance between attractants and deterrents to which the insects respond. Host plant ranking by moths and larvae of Chilo partellus Swinhoe (Lepidoptera: Crambidae), an important pest of cereals in Africa, was investigated, and whether eggs deposited on specific host plants yield larvae of particular host preferences. Trap plants are used in management of this pest. However, any ‘disagreement’ in host ranking between moths and larvae could potentially reduce effectiveness of trap crops as larvae emigrate to the main crop from the parent’s preferred trap plant. We also investigated whether host plant preference is influenced by the diet upon which larvae fed as part of an integrated assessment of the relationship between host plant selection and learning in C. partellus. Five host plants (all Poaceae) were used: maize (Zea mays L.), sorghum (Sorghum bicolor Moench), Napier grass (Pennisetum purpureum Schumach), and two varieties of signal grass [Brachiaria brizantha (A. Rich.) Stapf], viz., local (henceforth signal grass) and improved (‘Mulato’). In multiple choice tests, C. partellus female moths preferentially oviposited on Napier grass, followed by sorghum, maize, and signal grass, and least preferred ‘Mulato’. Larvae however equally orientated and settled on leaf cuts of maize, sorghum, signal grass, and Napier grass, but least preferred ‘Mulato’. Moreover, eggs from specific host plants did not yield larvae of particular host preferences. Furthermore, oviposition preference was not altered by the larval food. These results imply only a slight ‘disagreement’ in host ranking behaviour between moths and larvae, which is beneficial for trap cropping as larvae would not ‘reject’ the trap plant and appreciably disperse to the neighboring plants. Moreover, absence of larval learning behaviour indicates that regardless of the larval food C. partellus moths would still be attracted to the selected trap plant.     

Evaluation of Napier grass (Pennisetum purpureum) varieties for use as trap plants for the management of African stemborer (Busseola fusca) in a push–pull strategy

We evaluated eight Napier grass [Pennisetum purpureum Schumach (Poaceae)] varieties, used in various parts of eastern Africa as fodder, for their potential role as trap plants in the management of the African stemborer, Busseola fusca Füller (Lepidoptera: Noctuidae) through a push–pull strategy. Oviposition preference, larval orientation, settling, arrest and dispersal, feeding, mortality and survival, and development were determined for each of these varieties under laboratory and screen house conditions. Two‐choice tests showed that only two of the varieties tested (cv. Bana and cv. Uganda Hairless) were preferentially chosen by gravid female moths for oviposition over a susceptible maize variety, cv. Western Hybrid 502. Larval preference was, however, highly variable. Larval feeding by first instars on the maize leaves was more intense and significantly more than on leaves of all the Napier grass varieties evaluated. Food consumed and amounts assimilated by the third instars over a 24‐h period were not different among larvae fed on stems of maize and those fed on stems of the various Napier grass varieties. Larval survival was significantly lower on all the Napier grass varieties (below 3%) than on maize (about 44%). Similarly, larval development was about 2–3 weeks longer on majority of the Napier grass varieties. It was concluded that cv. Bana had potential for use as a trap plant in the management of B. fusca because it was more preferred by the moths for oviposition, equally preferred as maize by the larvae for orientation, settling, and arrest, and allowed minimal survival of the larvae. It can thus be used with such ‘push’ plants as Desmodium spp. (Fabaceae) in a ‘push–pull’ strategy, but the effectiveness of such a strategy would strictly depend on proper establishment and management of these companion plants.     

Proximate Composition of Freshwater Fish — Grass Carp,Puntius gonionotus and Tilapia

Summary The proximate analyses of freshwater fish — Grass carp (Ctenopharyngodon idellus), Puntius gonionotus and Tilapia (male hybrid) are compared. Dried meals from eviscerated freshwater fish contained 57.36–75.92% protein; 4.48–31.84% fat and 10.68–22.88% ash. The results indicate that Tilapia had a higher fat content than gonionotus and grass carp, the latter having the lowest fat content. The reverse is true for protein, ash and minerals like sodium, phosphorus and potassium. The composition of grass carp feeding on different diets, such as Hydrilla verticillata, Napier grass (Pennisetum purpureum) and tapioca leaves (Manihot utilissins) indicated only a slight difference in protein content but a significant difference in fat and ash. The tabulated results show that grass carp and goniotus were of higher nutritive value when compared to the Tilapia.     

Composition and nutritive value of some grasses, plants and aquatic weeds tested as diets

The composition and nutritive value of different diets, namely the aquatic weeds— Hydrilla verticillata (L.f.) Presl., Lemna gibbs (L.) Pistia stratiotes (L.), grasses—Guinea grass Panicum maximum Jacq., Napier grass Pennisetum purpureum Schum. and plants—sweet potato leaves Ipomaea batatas L. and tapioca leaves Manihot utilissimus Pohl. were determined and compared. The diets contained 13.5 to 30.48% protein; 1.89 to 9.60% fat; 3.94 to 23.13% ash; 5.42 to 26.70% fibre and 24.87 to 46.27% carbohydrate. Most of the minerals determined in the aquatic weeds were higher than those in grasses and plants. The feeding of Hydrilla , Napier grass and tapioca leaves to grass carp Ctenopharyngodon ideltus (V.) showed a different growth rate and the results obtained indicated the superiority of Hydrilla diet over Napier grass and tapioca leaves.     

柊叶和象草波式潜流人工湿地的对比研究

为探索柊叶和象草在人工湿地中的应用及其净化机理,该研究以柊叶和象草为人工湿地植物分别构建了波式潜流人工湿地系统,分析了柊叶和象草波式潜流人工湿地对生活污水中COD_(cr)、TN和TP的净化效果,观察了柊叶和象草两种植物在不同季节的生长状况。结果表明:经过15个月的连续运行,在表面水力负荷约0.3 m·d~(-1)的条件下,柊叶和象草波式潜流人工湿地平均去除率是COD_(cr)分别为66.1%和70.1%,TN分别为60.4%和63.7%,TP分别为74.1%和75.1%。两种植物生长良好,根系发达,象草的地上生物量是柊叶的2.1倍,地下生物量相当;冬季象草生长缓慢,柊叶部分叶片的四周干枯,但二者都不会枯亡。这说明两个人工湿地对COD_(cr)、TN和TP都具有较好的去除效果,但无显著性差异,柊叶和象草能明显提高潜流人工湿地的净化效果。     

Flight and oviposition behaviour of the African stem borer,Busseola fusca,on various host plant species

The African stem borer, Busseola fusca (Fuller) (Lepidoptera: Noctuidae), is an important pest of maize and sorghum in sub‐Saharan Africa. As in many other lepidopteran insects, the ability of B. fusca to recognize and colonize a variety of plants is based on the interaction between its sensory systems and the physical and chemical characteristics of its immediate environment. In this study, we tried to identify the behavioural steps of B. fusca leading to host selection and oviposition. Three Poaceae species commonly cultivated in Kenya for human consumption and animal forage were used in this study: the two most preferred hosts, maize (Zea mays L.) and sorghum [Sorghum bicolor (L.) Moench], and one non‐preferred host, Napier grass (Pennisetum purpureum Schumach). Wind tunnel observations revealed that volatiles produced by the different plant species did not appear to strongly influence the general orientation of B. fusca towards the plant, as similar behavioural steps were exhibited by the female moth regardless of the plant species involved. This indicated that the females were not able to recognize their preferred hosts from a distance. After landing, the female typically swept her ovipositor on the plant surface, simultaneously touching it with the tips of her antennae, and then oviposited. This behaviour was more frequently observed on maize and sorghum than on Napier grass, and indicated that both antennal and ovipositor receptors are used by the female moths to evaluate the plant surface before deciding to oviposit. As a result, the females laid more eggs on the two crops than on Napier grass. We conclude therefore that females recognized their preferred hosts only after landing. Tactile and contact‐chemoreception stimuli from the plants seemed to play a major role in oviposition decisions of B. fusca.     

Genome survey sequencing of purple elephant grass (<Emphasis Type="Italic">Pennisetum purpureum</Emphasis> Schum ‘Zise’) and identification of its SSR markers

Elephant grass (Pennisetum purpureum) is a perennial grass in the Poaceae family with high tolerance and one of the best forage plants. Despite its economic importance, the inheritance information of P. purpureum has remained largely unknown. To obtain the whole reference genome, we first conducted a genome survey of P. purpureum. Next-generation sequencing (NGS) was used to perform the de novo whole genome sequencing. As a result, the estimated genome size of elephant grass was 2.01 Gb, with 71.36% repetitive elements. The heterozygosity was 1.02%, which indicates a highly heterozygous genome. The retroelements (9.36%) were the most repetitive elements, followed by DNA transposons (3.66%). In the meantime, 83,706 high-quality genomic simple sequence repeat (SSR) markers, in which the greatest SSR unit length was 3, were developed. Thirty pairs of SSR markers were randomly selected to verify the efficiency and all of them yielded clear amplification products, among which 28 pairs (93.3%) of the primers showed polymorphism. The genome data obtained in this research provided a large amount of gene resources for further investigating Pennisetum species.     

Economic Competitiveness of Napier Grass in Irrigated and Non-irrigated Georgia Coastal Plain Cropping Systems

Interest and focus on development of renewable biofuels has been increasing over the past decade leading to the introduction of a wide cadre of renewable feedstocks. As a result, numerous perennial warm-season grasses have been introduced and management practices evaluated to determine their suitability as biofuel feedstocks. “Merkeron” napier grass (Pennisetum purpureum) plots were established in 2010 and harvested during crop years 2011 through 2015 adjacent to an on-going peanut (Arachis hypogaea L.), corn (Zea mays L.), and cotton (Gossypium hirsutum L.) cropping systems study conducted at the USDA/ARS Multi-crop Irrigation Research Farm in Shellman, GA (84 36 W, 30 44 N) on a Greenville fine sandy loam (fine, kaolinitic, thermic Rhodic Kandiudults). Napier grass was produced in both non-irrigated and two irrigated levels with different levels of nitrogen and potassium fertilizers. Peanut, corn, and cotton were produced in non-irrigated and full irrigation regimes. Breakeven prices for napier grass ranged from $65 to $84 Mg^?1 at variable and total costs. The breakeven napier grass price was estimated such that the net returns were equal between napier grass and peanut, cotton, corn cropping systems. At variable production cost, comparative breakeven napier grass prices for non-irrigated, 50% irrigated, and full irrigated regimes were $77, $117, and $112 Mg^?1, respectively. Napier grass did not compete economically against traditional irrigated cropping systems. Depending on traditional crop prices and bioenergy feed stock prices, napier grass could offer economic opportunities in non-irrigated production environments, riparian buffer zone edges, or non-cropped marginal production areas.     

Effect of wild grasses planted as border rows on stemborer infestations in maize in Uganda)

The presence of wild grasses in the vicinity of maize fields has been associated with reduced stemborer incidence on the maize crop. This study evaluated the impact of border rows with wild grasses on stemborer infestations and parasitism. Field trials were conducted in East and Central regional Agricultural Research Institutes in Uganda for three seasons. The four grass species planted as border rows included Pennisetum purpureum Schumach, Pennisetum polystachion (L.) Schult, Panicum maximum Jacq. and Sorghum arundinaceum (Desv.) Stapf. (Poaceae) were used. A pure maize stand without a grass border was planted as control. Busseola fusca Fuller (Lepidoptera: Noctuidae) and Chilo partellus Swinhoe (Lepidoptera: Crambidae) were the major stemborers found. Maize with P. purpureum and P. maximum borders reduced stemborer infestations and damage compared to the pure maize stand. The results were however, not consistent for all the three seasons. Yield per plot was higher in pure maize stand compared to plots with grass borders. The usefulness of this technique in stemborer management is discussed.     

Amaranthus caudatus subsp. mantegazzianus: A new host of ‘Candidatus Phytoplasma hispanicum’ (subgroup 16Sr XIII‐A)

In Argentina, amaranth is a promising crop due to high nutritional quality and ability to grow in a diversity of environments. In areas cultivated with amaranth, were observed plants exhibiting slow growth, deformed leaves, proliferation of shoots and malformed lateral panicles. Field survey revealed up to 96% disease incidence and 92% of the seeds collected from mother plants produced diseased seedlings. A phytoplasma was detected in association with seedlings and adult plants using nested PCR assays. Molecular identification by computer‐simulated RFLP and phylogenetic analysis evidenced the occurrence of a ‘Candidatus Phytoplasma hispanicum’‐related strain, affiliated with 16SrXIII‐A subgroup. The findings implicate amaranth as a new host for this subgroup and as a potential reservoir of the pathogen for other cultivated species. In addition, to the best of our knowledge, this study reports for the first time the presence of 16SrXIII‐A phytoplasma in Argentina and in South America. Furthermore, transmission assays pointed that naturally infected seed is an important vehicle of dissemination of the pathogen, threatening the expansion of the crop for new geographical areas.     

Mixed Infection and Natural Spread of ‘Candidatus Phytoplasma asteris’ and Mungbean Yellow Mosaic India Virus Affecting Soya Bean Crop in India

Suspected phytoplasma and virus‐like symptoms of little leaf, yellow mosaic and witches’ broom were recorded on soya bean and two weed species (Digitaria sanguinalis and Parthenium hysterophorus), at experimental fields of Indian Agricultural Research Institute, New Delhi, India, in August–September 2013. The phytoplasma aetiology was confirmed in symptomatic soya bean and both the weed species by direct and nested PCR assays with phytoplasma‐specific universal primer pairs (P1/P6 and R16F2n/R16R2n). One major leafhopper species viz. Empoasca motti Pruthi feeding on symptomatic soya bean plants was also found phytoplasma positive in nested PCR assays. Sequencing BLASTn search analysis and phylogenetic analysis revealed that 16Sr DNA sequences of phytoplasma isolates of soya bean, weeds and leafhoppers had 99% sequence identity among themselves and were related to strains of ‘Candidatus Phytoplasma asteris’. PCR assays with Mungbean yellow mosaic India virus (MYMIV) coat‐protein‐specific primers yielded an amplicon of approximately 770 bp both from symptomatic soya bean and from whiteflies (Bemisia tabaci) feeding on soya bean, confirmed the presence of MYMIV in soya bean and whitefly. Hence, this study suggested the mixed infection of MYMIV and ‘Ca. P. asteris’ with soya bean yellow leaf and witches’ broom syndrome. The two weed species (D. sanguinalis and P. hysterophorus) were recorded as putative alternative hosts for ‘Ca. P. asteris’ soya bean Indian strain. However, the leafhopper E. motti was recorded as putative vector for the identified soya bean phytoplasma isolate, and the whitefly (B. tabaci) was identified as vector of MYMIV which belonged to Asia‐II‐1 genotype.     

Biochemical and molecular analysis of plants derived from embryogenic tissue cultures of napier grass (Pennisetum purpureum K. Schum)

Summary We have investigated the extent of biochemical and molecular variation in 63 plants of napier grass (Pennisetum purpureum K. Schum.) regenerated from 3- to 24-week-old embryogenic callus cultures. The calli were derived from cultured basal segments of young leaves and immature inflorescences obtained from a single fieldgrown donor plant. The entire population was analyzed for the activity of 14 isozyme systems, but no qualitative variation was found at any of the loci examined. Similarly, no restriction fragment length polymorphisms (RFLPs) were detected in the mitochondrial, plastid and nuclear genomes in a representative sample of regenerated plants. Our results confirm earlier reports of the genetic uniformity of plants derived from somatic embryos and highlight their value both for clonal propagation and for genetic transformation.     

Occurrence and identification of aster yellows related phytoplasma in Gladiolus in Poland

In 1996–1998 on Gladiolus plants cultivated in Poland severe symptoms were observed. The symptoms included chlorosis of the youngest leaves, yellowing and malformation of flower spices, flower discoloration and virescence. The affected corms kept in cold storage developed premature multiple sprouts weak and pale in color. Their root formation was strongly inhibited. Electron microscopy examination of the ultra-thin sections of the leaves and roots of diseased plants showed necrosis and collapsing of sieve tubes and companion cells, reduction of phloem and xylem strands as well as decrease of the number and diameter of xylem vessels. Numerous polymorphic bodies were observed in the phloem and parenchyma cells of affected gladioli. PCR amplification using universal phytoplasma primers rU3 and fU5 directed to ribosomal sequences and RFLP analysis of the amplified rDNA were used to identify the phytoplasma causing yellow disease in Poland. Specific product of about 880 bp was obtained, providing evidence of phytoplasma infection. RFLP analysis of the PCR product done with restriction enzyme AluI showed that the diseased gladioli were infected by phytoplasma very similar or identical with American aster yellows phytoplasma.     

A new host record for Candidatus Phytoplasma cynodontis (16Sr XIV-A) associated with phyllody and fasciation of linseed (Linum usitatissimum) from India

Linseed commonly called as flaxseed (Linum usitatissimum Linn.) is an important oilseed crop cultivated widely in Northern parts of Karnataka. During, 2019 (January–February), a characteristic disease was noticed with symptoms that resembled phytoplasma or like disease symptoms. The incidence was ranged from 6·5 to 16·5% in the experimental station of Raichur Agricultural University. The typical symptoms observed were virescence of floral parts, fasciation of the inflorescence axis, phyllody, stunted and flattened stem with reduced leaves. Symptomatic and healthy samples were collected and processed for molecular detection of phytoplasma. Total DNA was isolated from four infected plants and two healthy plants. The 16S rDNA region was amplified using P1/P7 followed by R16F2n/R16R2 primer pair which showed the amplification of expected amplicon size from all four infected samples. Furthermore, the SecA gene was amplified using SecA1/SecA3 primers. The PCR amplified products were subjected for direct sequencing from both directions and the consensus sequences were obtained and nBLAST search analysis revealed that the 16Sr RNA and SecA sequences were sharing maximum similarity (100%) with the reference sequence of Ca. P. cynodontis. The sequences were analysed phylogenetically by constructing a Phylogram independently by NJ method along with reference sequence of 16S rRNA region and SecA region retrieved from GenBank database showed that the phytoplasma sequence from linseed phyllody of the present study placed in a distinct clade along with reference sequence of “Ca. P. cynodontis” thus confirming the identity phylogenetically. Furthermore, iPhyClassifier and virtual RFLP proved that the phytoplasma belonged to 16SrXIV (subgroup A) phytoplasma. Previously linseed is known to be associated with 16SrII-D phytoplasma but the association of the 16SrXIV-A group of phytoplasma is not reported so far. Therefore, this is the new host record for Ca. P. cynodontis (16SrXIV-A) phytoplasma associated with linseed stem fasciation, phyllody from India.