Aphid Inoculation Research Articles

Transcriptomic Analysis Revealed That Low-Density Aphid Infestation Temporarily Changes Photosynthesis and Disease Resistance but Persistently Promotes Insect Resistance in Poplar Leaves

Insect infestations cause substantial changes in the leaves they attack, but the effects of insect infestation on photosynthesis and gene expression in the whole infestation period are rarely reported. In this study, the effects of poplar aphid (Chaitophorus populialbae Boyer de Fonscoloube) on photosynthetic rate and gene expression of Populus alba var. pyramidalis were studied, from 2 to 36 days after low-density aphid inoculation (DAI). The results revealed a dynamic change of photosynthesis in poplar after aphid infestation: compensatory increased at 8 DAI, was inhibited at 17 DAI, but repaired after 21 DAI. Aphid infestation promoted the expression of the majority of differentially expressed genes (DEGs) at 4, 6, 8, and 21 DAI. The DEGs were enriched into a plant–pathogen interaction, plant hormone signal transduction, and MAPK signaling pathway, suggesting a significant but transient resistance to disease or other stresses. Photosynthesis-related DEGs were downregulated at 8 DAI, which might result in photosynthetic inhibition at 17 DAI. The expression of flavonoid biosynthesis-related DEGs dynamic changed from all downregulated at 2 DAI to almost all upregulated at 21 DAI, suggesting a persistent resistance to insect infestation. This study reveals dynamic changes in photosynthesis, resistance to pathogens, and insects in poplar leaves after aphid infestation.

RNA-dependent RNA polymerase could extend the lasting validity period of exogenous dsRNA.

Previous studies have found that pesticide double-stranded (ds)RNA usually has a long-lasting validity period in plants. However, it is uncertain if any factors in plants could extend dsRNA duration. It has been reported that RNA-dependent RNA polymerases (RdRP) in plants and some other eukaryotes could catalyze RNA amplification and be involved in RNAi (interference). Thus, this study evaluated the effect of RdRP on the tissue content, activity, and duration of exogenous dsRNA. We found that RdRP knockdown in Arabidopsis thaliana had no significant effect on tissue contents of reporter dsRNA parent molecules (8.91% reduction), but it caused significant decrease in the tissue contents of derived short fragments of 200, 120 and 59 bp tested (51.22%, 52.83% and 59.35%, respectively). Aphid inoculation tests showed that the same dose of insecticidal dsAgZFP exhibited a significantly lower lethal effect (mortality 58.8%) in the plants with RdRP knockdown than in the control plants with normal RdRP (86.0%). For Caenorhabditis elegans, the worms treated simultaneously with dsRdRP and reporter dsRNA had similar body contents to reporter dsRNA parent molecules and its long-fragment derivative (200 bp) as the control (1.28- and 1.07-fold greater, respectively). However, 120- and 59-bp short-fragment derivatives were significantly reduced by 28.78% and 59.84%, respectively, which also diminished faster in the descendants. We conclude that RdRP could significantly enhance the tissue content of dsRNA derivatives by catalyzing amplification, thus improving dsRNA activity and extending its lasting validity period. Otherwise, RNAi by exogenous dsRNA was proven to be noninheritable in A.thaliana. This work confirmed the merit of dsRNA as a plant protectant. © 2022 Society of Chemical Industry.

Novel Sources of Turnip Yellows Virus Resistance in Brassica and Impacts of Temperature on Their Durability.

Turnip yellows virus (TuYV; family Solemoviridae, genus Polerovirus) is the most widespread and economically damaging virus of canola (Brassica napus L.) production in Australia. However, no Australian commercial seed companies market TuYV-resistant canola cultivars, and little information is available on the susceptibility of those available. To identify potential sources of TuYV resistance, 100 B. napus accessions from the ERANET ASSYST diversity set were screened in the field and five of these were selected for further phenotyping via aphid inoculation. Furthermore, 43 Australian canola cultivars, six B. napus genotypes with previously reported resistance, and 33 B. oleracea and B. rapa cultivars were phenotyped. All Australian cultivars were susceptible except for 'ATR Stingray'. Stronger resistance to systemic TuYV infection (IR) was identified in diversity set accessions 'Liraspa-A', 'SWU Chinese 3', and 'SWU Chinese 5'. As indicated by lower relative enzyme-linked immunosorbent assay absorbance values (R-E405) in infected plants, resistance to TuYV accumulation (AR) often accompanied IR. Moderate IR was identified in four B. oleracea cultivars and one B. rapa cultivar. Very strong AR was identified in four B. oleracea cultivars and AR of some degree was common across many cultivars of this species tested. The impact of temperature during the inoculation access period or post-inoculation incubation on the resistance identified was examined. Infection rates were significantly higher in resistant B. napus genotypes when inoculated at 16°C than at 26°C, suggesting an increase in aphid transmission efficiency. IR in B. napus genotypes was strong when incubated at 16°C, but weakened at elevated temperatures with almost total breakdown in most genotypes at 30°C. However, infected plants of B. napus and B. oleracea genotypes with AR maintained lower R-E405 values than susceptible controls at all temperatures tested. Novel sources of resistance identified in this study offer potential as breeding material in Australia and abroad.

Honeydew Deposition by the Giant Willow Aphid (Tuberolachnus salignus) Affects Soil Biota and Soil Biochemical Properties.

Infestation of willow plants by the giant willow aphid Tuberolachnus salignus (Hemiptera: Aphididae) is associated with copious deposition of sugar-rich honeydew under the plant canopy. We explored the effect of aphid honeydew on the soil biota and biochemical indicators in a two-year field trial. Soil samples from under aphid-infested and control willow trees, as well as samples from black sooty mould spots under the aphid-infested willows were compared; soil samples before aphid inoculation were used as a baseline. The honeydew deposition had a positive effect on the total soil carbon (C), but not on the total soil nitrogen content or soil pH. Microbial biomass C, basal respiration, number of yeast colony forming units, and the geometric mean of activities for six enzymes were significantly higher in honeydew-affected soils than in the control treatment on both years. The honeydew deposition also increased soil meso-fauna abundance, especially in the black sooty mould spots. The soil biochemical properties, which differed before and after aphid infestation, showed considerable overlap between the first and second year post-infestation. The results highlight the cascading effects of T. salignus on soil biological activity and the importance of using a multitrophic approach to explore similar scenarios.

Gamma Radiation and In Vitro Induced Banana Bunchy Top Virus (BBTV) Resistant Mutant Lines of Banana cv ‘Lakatan’ (Musa sp., AA)

Gamma irradiation coupled with in vitro technology was used to develop BBTV resistance in banana cv ‘Lakatan.’ Ten (10) resistant lines were selected after several generations of evaluation and selection. The selected lines (M1V3–M1V4) showed low disease incidence in the field under high disease pressure and, likewise, low disease incidence with aphid inoculation of the virus. Further disease evaluation (M1V4–M1V5) on these lines consistently showed low BBTV disease incidence (11.62–28.57%) 30 mo from planting (MAP). The genetic variability in morpho-agronomic traits derived from SHAN cluster analysis grouped the selected lines into four major clusters for qualitative traits at 0.636 coefficient of similarity and three clusters for quantitative traits at 0.11 Euclidian distance coefficient. Selections based on agronomic traits showed significantly earlier flowering in three mutant lines and shorter stature in two mutant lines. Short strand repeat (SSR) analysis using 11 primers detected a high level of polymorphism in mutant lines. Mutant lines were differentiated from the ‘Lakatan’ control by the absence of one or few alleles in mutant lines with four primers and/or addition of one or few alleles in mutant lines with two primers. SSR analysis revealed genetic differences among mutant lines and between mutant lines and ‘Lakatan' control. The results of the study further affirmed stable BBTV resistance in advanced generation evaluation (M1V4M1V5). Five out of ten resistant lines were selected for further evaluation in multi-location field trials as a requirement for registration and release of new BBTV resistant mutant variety of ‘Lakatan.’

Elicitors and defense gene induction in plants with altered lignin compositions.

A reduction in the lignin content in transgenic plants induces the ectopic expression of defense genes, but the importance of altered lignin composition in such phenomena remains unclear. Two Arabidopsis lines with similar lignin contents, but strikingly different lignin compositions, exhibited different quantitative and qualitative transcriptional responses. Plants with lignin composed primarily of guaiacyl units overexpressed genes responsive to oomycete and bacterial pathogen attack, whereas plants with lignin composed primarily of syringyl units expressed a far greater number of defense genes, including some associated with cis-jasmone-mediated responses to aphids; these plants exhibited altered responsiveness to bacterial and aphid inoculation. Several of the defense genes were differentially induced by water-soluble extracts from cell walls of plants of the two lines. Glycome profiling, fractionation and enzymatic digestion studies indicated that the different lignin compositions led to differential extractability of a range of heterogeneous oligosaccharide epitopes, with elicitor activity originating from different cell wall polymers. Alteration of lignin composition affects interactions with plant cell wall matrix polysaccharides to alter the sequestration of multiple latent defense signal molecules with an impact on biotic stress responses.

Soil microbial species loss affects plant biomass and survival of an introduced bacterial strain, but not inducible plant defences.

Plant growth-promoting rhizobacteria (PGPR) strains can influence plant-insect interactions. However, little is known about the effect of changes in the soil bacterial community in general and especially the loss of rare soil microbes on these interactions. Here, the influence of rare soil microbe reduction on induced systemic resistance (ISR) in a wild ecotype of Arabidopsis thaliana against the aphid Myzus persicae was investigated. To create a gradient of microbial abundances, soil was inoculated with a serial dilution of a microbial community and responses of Arabidopsis plants that originated from the same site as the soil microbes were tested. Plant biomass, transcription of genes involved in plant defences, and insect performance were measured. In addition, the effects of the PGPR strain Pseudomonas fluorescens SS101 on plant and insect performance were tested under the influence of the various soil dilution treatments. Plant biomass showed a hump-shaped relationship with soil microbial community dilution, independent of aphid or Pseudomonas treatments. Both aphid infestation and inoculation with Pseudomonas reduced plant biomass, and led to downregulation of PR1 (salicylic acid-responsive gene) and CYP79B3 (involved in synthesis of glucosinolates). Aphid performance and gene transcription were unaffected by soil dilution. Neither the loss of rare microbial species, as caused by soil dilution, nor Pseudomonas affect the resistance of A. thaliana against M. persicae. However, both Pseudomonas survival and plant biomass respond to rare species loss. Thus, loss of rare soil microbial species can have a significant impact on both above- and below-ground organisms.

Experimental transmission efficiency of Plum pox virus T by Myzus persicae Sulzer (Homoptera: Aphididae)

Plum pox virus (PPV), which is a major threat to stone fruit trees, is transmitted by various aphid species. Of the nine PPV strains, PPV-T is a unique strain found only in Turkey. In this study, PPV-T was experimentally transmitted to Nicotiana benthamiana Domin by using Myzus persicae with the unrestricted probing method (UPM) and the controlled acquisition access period (CAAP) method. Three weeks after aphid inoculation, PPV symptoms were observed on some inoculated tobacco plants. When test plants were analysed by DAS-ELISA and real-time RT-PCR, only the symptomatic plants were found positive for PPV, and the infection rate was found to be 14.29% for both methods by using UPM, whereas it was 5.40% in CAAP only by real-time RT-PCR. Furthermore, it was revealed that PPV-T was transmitted more efficiently to N. benthamiana by M. persicae by using the UPM method.

Elevated CO2 impacts bell pepper growth with consequences to Myzus persicae life history, feeding behaviour and virus transmission ability.

Increasing atmospheric carbon dioxide (CO2) impacts plant growth and metabolism. Indirectly, the performance and feeding of insects is affected by plant nutritional quality and resistance traits. Life history and feeding behaviour of Myzus persicae were studied on pepper plants under ambient (aCO2, 400 ppm) or elevated CO2 (eCO2, 650 ppm), as well as the direct impact on plant growth and leaf chemistry. Plant parameters were significantly altered by eCO2 with a negative impact on aphid’s life history. Their pre-reproductive period was 11% longer and fecundity decreased by 37%. Peppers fixed significantly less nitrogen, which explains the poor aphid performance. Plants were taller and had higher biomass and canopy temperature. There was decreased aphid salivation into sieve elements, but no differences in phloem ingestion, indicating that the diminished fitness could be due to poorer tissue quality and unfavourable C:N balance, and that eCO2 was not a factor impeding feeding. Aphid ability to transmit Cucumber mosaic virus (CMV) was studied by exposing source and receptor plants to ambient (427 ppm) or elevated (612 ppm) CO2 before or after virus inoculation. A two-fold decrease on transmission was observed when receptor plants were exposed to eCO2 before aphid inoculation when compared to aCO2.

Aphid Transmission of the Ontario Isolate of Plum Pox Virus.

Utilization of timed virus acquisition access probes in studies of plum pox virus (PPV) transmission by aphids demonstrated that endemic species transmitted the virus readily from plum, Prunus domestica (L.) Batsch; peach, P. persica (L.); or dwarf flowering almond, P. glandulosa Thunberg., to peach seedlings. The green peach aphid, Myzus persicae (Sulzer), was shown to be the most efficient vector. Acquisition of virus by green peach aphids from infected peach leaves resulted in 18-28% infected peach seedlings, while aphids previously fed on infected leaves of plum transferred virus to 36% of peach seedlings. Although the spirea aphid, Aphis spiraecola (Patch), was a less efficient vector than M. persicae it is perhaps more important for the spread of PPV due to its greater abundance and occurrence earlier in the season when peach trees are thought to be more susceptible to infection. Virus transmission rates varied depending on the virus source and healthy test plant species. In contrast to many previous studies, aphid inoculation of the experimental host Nicotiana benthamiana Domin occurred at a low rate, never exceeding 4%. Acquisition of PPV by M. persicae from infected peach fruit was greatly reduced compared with acquisition from leaves. The results of this research indicate that the Ontario isolate of PPV-D is readily transmissible by aphids to peach and natural spread of the virus needs to be considered in future management or eradication programs.

Detection and partial genome sequence of a new umbra-like virus of papaya discovered in Ecuador

Sequencing of cDNA synthesized from double-stranded RNA (dsRNA) from papaya leaves revealed the presence of a putative new virus with homology to genomes of several members of the genus Umbravirus. A wide survey showed that the virus was present in two out of three papaya producing provinces in Ecuador included in this study. The virus was found in up to 31 % of symptomatic trees co-infected with Papaya ringspot virus (PRSV) compared to 2 % in symptomless PRSV-free plants. Mechanical and aphid inoculations failed to transmit the virus under controlled conditions. The virus was not transmitted through seeds. The presence of a luteovirus, commonly associated with umbraviruses, was not confirmed in plants infected with the new virus using universal luteovirus primers.

First Report of Bean common mosaic virus Infecting Mungbean (Vigna radiata) in China.

Bean common mosaic virus (BCMV) is a member of the genus Potyvirus and one of the numerous viruses that can infect Phaseolus vulgaris. In May of 2013, we planted more than 100 varieties of mungbean in fields and a greenhouse of Nanjing. Mungbean (Vigna radiata (Linn.) Wilczek.) with leaves displaying mosaic and shrinkage typical of viral infection was observed in a greenhouse and a field in Nanjing. About 60% of the varieties can be infected. The symptoms in some sources from Southeast Asian countries and wild germplasm are heavier, while the symptoms are lighter on the local varieties. It can be transmitted to mungbean via aphid or mechanical inoculation, or by seeds. The infected leaves were collected for electron micrograph analysis. Pinwheel inclusion and filamentous virus particles were observed, indicating a Potyvirus infection. To confirm the presence of Potyvirus infection, total RNA was extracted from plants from the greenhouse and field, and RT-PCR was performed using universal Potyvirus primers (Sprimer (+) 5'-GGXAAYAAYAGYGGXCAZCC-3'; X=A, G, C or T, Y=T or C, Z=A or G); M4T (5'-GTTTTCCCAGTCACGAC(T)-3'), which amplify a region of the 3' fragment of the RNA-dependent RNA polymerase of potyviruses (1). The 596-bp sequence was found to be 95% identical to the BCMV isolate HB (GenBank Accession No. KC478389.1). To confirm the presence of BCMV, three leaf samples were randomly collected and all were determined to be positive when subjected to ELISA using BCMV-specific antibodies. The virus infecting mungbean was identified as BCMV and the strain was named BCMV-JAAS (KJ866945). Using gene-specific primers (BCMV-cp-F: 5'-CAAAAGGACAAGGATTGAGGA, BCMV-cp-R: 3'-ACAACAAACATTGCCGTAGC) for the reported coat protein gene in BCMV, a 1,080-bp gene fragment was amplified from the total RNA of the isolate, and subsequent sequence analysis indicated that an 862-bp region contained the complete cp gene that encodes a 228 amino acid protein. The nucleotide sequences of the cp gene from the isolate shared 96% homology with the reported BCMV-HB. The phylogenetic trees based on the CP gene show that BCMV-JAAS (KJ866945) was most closely related to other Chinese BCMV isolates (KF439722.1 and AJ132145.1) followed by Azuki mosaic virus (AB012663.1) and Peanut stripe virus (U34972.1). These results indicate that the virus associated with the mosaic disease in mungbean is an isolate of BCMV. To our knowledge, this is the first report of BCMV infecting mungbean in China. BCMV affects a wide range of legume crops and can spread rapidly, causing serious harm. The discovery could effectively control BCMV and characterize the prevalent BCMV strains. Research utilizing whole-genome sequencing of the mungbean isolate is continuing and is currently being expanded to characterize the genetic diversity of the virus, assisting in the study of the evolution of the virus. Reference: (1) J. Chen and J. P. Chen. Chin. J. Virol. 18:372, 2002.

First Report of Clover yellow vein virus on White Clover (Trifolium repens) in South Korea.

White clover (Trifolium repens L.) is a herbaceous, perennial plant that has become one of the most widely distributed legumes in the world. It is extensively used in grass-legume pastures, but also has the potential to invade agricultural lands and natural ecosystems. White clover is a well-known natural host for Alfalfa mosaic virus (AMV), Clover yellow vein virus (ClYVV), Soybean dwarf virus (SbDV), Beet western virus (BWYV), Tomato spotted wilt virus (TSWV), Zucchini yellow mosaic virus (ZYMV), etc (1). In July 2013, during a survey to determine the presence of different viruses infecting weed plants in South Korea, three white clover leaf samples showing yellow mosaic symptoms were collected from Taean County, South Chungcheong Do Province, South Korea. In order to identify the infecting virus, total RNA from three leaf samples was extracted using the Tri-reagent (MRC Reagent, Inc., OH) as described by the manufacturer, and was applied to the large-scale oligonucleotide (LSON) chip (3), wherein probes specific to a ClYVV isolate produced a positive reaction. All three samples tested were positive for ClYVV. To confirm this result, ClYVV-specific primers were designed using the sequences of four ClYVV isolates from NCBI (GenBank Accession Nos. AF185959, AF203536, DQ333346, and NC003536). Total RNA was extracted from symptomatic white clover samples using Easy-Spin Total RNA Extraction Kit (iNtRon, Daejeon, Korea) and used as template for RT-PCR. The positive control RNA was used from ClYVV GM isolate (KF975894) and negative control RNA used symptomless white clover plants. The ClYVV coat protein (CP) gene was amplified by RT-PCR using the specific primer pairs ClYVV-CP-F / ClYVV-CP-R (5'-CAAGAGCAGCACGATGAG-3' and 5'-CTCGCTCTATAAAGATCAGAT-3'). DNA fragments of the expected size (1,042 bp) were obtained from the white clover Korea isolate (AB930132), and the PCR product was cloned into a T&A cloning vector (RBC Bioscience, Taipei, Taiwan) and sequenced directly in both directions. BLAST analyses of the nucleotide sequence CP gene fragments revealed the highest identity with 98% with other ClYVV isolates (AF203536). To determine the experimental host range of the ClYVV Korea isolate, we inoculated five species (Chenopodium amaranticolor, C. quinoa, Nicotiana clevelandii, N. benthamiana, and Trifolium repens) in three families using this isolate. All test plants were mechanically inoculated with 0.1 M phosphate buffered saline (Takara, Tokyo, Japan). Each test plant was inoculated nine times and grown in a greenhouse maintained at 27 to 33°C. Necrotic local lesions were produced on inoculated leaves of C. amaranticolor, C. quinoa, and N. clevelandii 4 to 6 days post-inoculation. After 10 to 14 days, C. amaranticolor and C. quinoa showed systemic chlorotic spot symptoms, and N. clevelandii, N. benthamiana, and T. repens showed chlorotic spot, mild mosaic, and mosaic in the upper leaves, respectively. Up to now, in South Korea, ClYVV has been detected in gladiolus (Gladiolus gandavensis) (3) and soybean (Glycine max) (4). ClYVV can be easily transmitted by insect, aphid, or mechanical inoculation and has a host range including tobacco, soybean, etc. The presence of ClYVV could become an important threat to crop production in South Korea. To our knowledge, this is the first report of a ClYVV infection of the white clover plant in South Korea.

Plant–rhizobia interactions alter aphid honeydew composition

Both above- and below-ground interspecific interactions contribute to ecosystem functioning in terrestrial systems, and the integration of below- and above-ground interactions is crucial for deepening our knowledge of nutrient cycling and community dynamics in terrestrial ecosystems. The present study explored the effects of plant–microbe interactions on aphid honeydew quality and quantity and important factors mediating ant–aphid mutualisms and below-ground nutrient dynamics. Soybean aphids (Aphis glycines) were inoculated onto two closely related strains of soybean plants: a nodulating strain that associates with rhizobia and a non-nodulating strain that does not harbor any nitrogen-fixing bacteria. As expected, prior to aphid inoculation, nodulating plants were significantly taller and had more leaves than non-nodulating plants. Aphids feeding on nodulating strains were found to reach slightly larger colony sizes and produce honeydew with significantly different sugar profiles than those feeding on non-nodulating plants. The honeydew collected from aphid colonies feeding on nodulating plants contained 160 % more total sugars than honeydew collected from colonies feeding on non-nodulating plants, but there was no difference in total amino acid-N content in honeydew from colonies feeding on the different plant strains. We discuss the implications of honeydew composition for nutrient cycling and community dynamics and suggest areas of future research to elucidate the consequences of altered aphid honeydew composition on ecosystem properties.

Biological studies of three caulimoviruses associated with dahlia (Dahlia variabilis)

Two distinct caulimoviruses, Dahlia mosaic virus (DMV) and Dahlia common mosaic virus (DCMV), were found to be associated with dahlia mosaic disease. Additionally, an endogenous caulimovirus, Dahlia variabilis endogenous plant pararetroviral sequence (DvEPRS), was found to be present in many cultivated and wild Dahlia spp. Biological properties including mechanical and aphid transmission were investigated for DMV, DCMV and DvEPRS using Verbesina encelioides (crownbeard) as an experimental host. DMV and DCMV were transmitted by Myzus persicae at the rate of 50 and 73%, respectively. DMV and DCMV were mechanically transmissible to V. encelioides and the two effective buffers for mechanical transmission were phosphate buffer (pH 7.0) with nicotinic acid (1%) and sodium bisulfite (5%). Mechanical transmission rates of 58% and 36% were obtained for DMV and DCMV, respectively. DvEPRS was not transmissible by either aphids or mechanical inoculation. Availability of aphid and mechanical inoculation protocols for DMV and DCMV would facilitate further biological studies on caulimoviruses associated with Dahlia.

The wild type of Momordica charantia is not infected by potyviruses that cause disease in papaya and cucurbit crops

In the present work, the susceptibility of wild and domesticated plants of Momordica charantia to viruses from the genus Potyvirus that cause diseases in papaya (Carica papaya) and cucurbit crops was evaluated. The plants were subjected to experimental and natural infection with Papaya ringspot virus types P and W (PRSV-P and PRSV-W) and Zucchini yellow mosaic virus (ZYMV). None of the potyviruses infected the wild type of M. charantia through mechanical or aphid inoculation or under field exposition, whereas the domesticated type was only infected by isolates of ZYMV. In addition, both wild and domesticated types of M. charantia were not infected in natural conditions by an isolate of Zucchini lethal chlorosis virus, genus Tospovirus, transmitted by Frankliniella zucchini. These data clearly indicated that this wild type of M. charantia does not seem to have a role in the epidemiology of the diseases caused by these three potyviruses in Brazil.

Transgenic banana plants expressing small interfering RNAs targeted against viral replication initiation gene display high-level resistance to banana bunchy top virus infection

The banana aphid-transmitted Banana bunchy top virus (BBTV) is the most destructive viral pathogen of bananas and plantains worldwide. Lack of natural sources of resistance to BBTV has necessitated the exploitation of proven transgenic technologies for obtaining BBTV-resistant banana cultivars. In this study, we have explored the concept of using intron-hairpin-RNA (ihpRNA) transcripts corresponding to viral master replication initiation protein (Rep) to generate BBTV-resistant transgenic banana plants. Two ihpRNA constructs namely ihpRNA-Rep and ihpRNA-ProRep generated using Rep full coding sequence or Rep partial coding sequence together with its 5' upstream regulatory region, respectively, and castor bean catalase intron were successfully transformed into banana embryogenic cells. ihpRNA-Rep- and ihpRNA-ProRep-derived transgenic banana plants, selected based on preliminary screening for efficient reporter gene expression, were completely resistant to BBTV infection as indicated by the absence of disease symptoms after 6 months of viruliferous aphid inoculation. The resistance to BBTV infection was also evident by the inability to detect cDNAs coding for viral coat protein, movement protein and Rep protein by RT-PCR from inoculated transgenic leaf extracts. Southern analysis of the two groups of transgenics showed that ihpRNA transgene was stably integrated into the banana genome. The detection of small interfering RNAs (siRNAs) derived from the ihpRNA transgene sequence in transformed BBTV-resistant plants positively established RNA interference as the mechanism underlying the observed resistance to BBTV. Efficient screening of optimal transformants in this vegetatively propagated non-segregating fruit crop ensured that all the transgenic plants assayed were resistant to BBTV infection.

Effect of cucumber mosaic virus infection on morphology, yield and phenolic contents of tomato

Ten tomato genotypes were screened for their resistance against cucumber mosaic virus (CMV) and its vector Myzus persicae under natural infection in field, using aphids M. persicae under net-house and mechanical inoculation under greenhouse. Large differences were observed among genotypes for infection percentage (IP) and severity index (SI) among the testing methods used. All genotypes showing tolerance to CMV in the field or through aphid inoculation, however, become susceptible and highly susceptible after mechanical inoculation. All the test genotypes also showed susceptibility to the aphid M. persicae population. Plants inoculated with CMV showed substantial decrease in yield and yield-contributing parameters which varied with cultivars that probably depended upon its genetic make up. All the test genotypes exhibited 0.97–30.19% decrease in plant height, 11.47–52.65% decrease in root length, 46.56–95.56% decrease in fresh plant weight, 65.78–92.84% decrease in root fresh weight, 19.97–87.65% decrease in the dry weight of plants, 75.63–95.43% decrease in dry root weight, 69.51–95.65% reduction in the number of fruits and 89.04–99.89% decrease in yield per plants. After 15 days of inoculation, the quantitative analysis using double beam spectrophotometer showed an increase in total phenolics in CMV-inoculated plants as compared to un-inoculated plants among genotypes. Similarly the thin layer chromatography (TLC) on silica gel G indicated that the number of phenolic compounds was increased in most of the inoculated genotypes while in others they were either decreased or remained same.

Within plant distribution of Potato Virus Y in hairy nightshade ( Solanum sarrachoides): An inoculum source affecting PVY aphid transmission

Potato virus Y (PVY) is vectored by several potato-colonizing and non-colonizing aphid species in a non-persistent manner and has a wide host range. It occurs naturally in several plant families. Myzus persicae and Macrosiphum euphorbiae are the most efficient potato-colonizing aphid vectors of PVY. Rhopalosiphum padi, a cereal aphid that migrates in large numbers through potato fields during the middle of the growing season, does not colonize potato plants but can transmit PVY. Hairy nightshade, Solanum sarrachoides, a prevalent annual solanaceous weed in the Pacific Northwest (PNW) of the United States, is an alternative host for PVY and a preferred host for M. persicae and M. euphorbiae. Hence, hairy nightshade plants might play an important role as an inoculum source in the epidemiology of PVY. We looked at titre accumulation and distribution of PVY O, PVY N:O and PVY NTN in S. sarrachoides and potato after aphid inoculation with M. persicae and studied the transmission of PVY O and PVY NTN, by M. persicae, M. euphorbiae and R. padi from hairy nightshade to potato plants. Virus titre at different positions on the plant was similar in S. sarrachoides and potato plants with strains PVY O and PVY N:O. Titres of PVY NTN were similar in S. sarrachoides and potato but differences in titre were observed at different positions within the plant depending on the plant phenology. Percentage transmission of PVY NTN by M. persicae and M. euphorbiae was twice as high (46 and 34%, respectively) from hairy nightshade to potato than from potato to potato (20 and 14%). Percentage transmission of PVY O by M. persicae and M. euphorbiae was not affected by the inoculum source. No effect of the inoculum source was observed in the transmission of either PVY strain by R. padi. These results show that hairy nightshade may be an equal or better virus reservoir than potato and thus, important in the epidemiology of PVY.

Genetics of corn leaf aphid(Rhopalosiphum maidis)resistance in barley

A study was undertaken to determine the genetics of corn leaf aphid (CLA) resistance in barley under controlled conditions with artificial inoculation at adult plant stage. Inheritance of CLA resistance was investigated in five resistant barley genotypes (EB921, EB2507, Manjula, DL529 and K144) in crosses with susceptible parent Alfa93 in F1, F2, F3 and backcross (BCF1) generations. The aphid inoculation was done using the brush method as well as the detached leaf method. Individual plants were classified in resistant, moderately resistant, susceptible and highly susceptible categories base on number per shoot as well as multiplication of CLA on any of the fresh, young leaf. The plants scored as resistant or moderately resistant were observed twice more at 10 days interval to confirm their reaction. Resistance was governed by a single dominant gene in EB921, DL529 and K144, while it was monogenic recessive in Manjula and EB2507. These diverse sources may be used in breeding for CLA resistance in barley im...