Structural and functional characterization of genes PYL-PP2C-SnRK2s in the ABA signalling pathway of Cucurbita pepo.

BACKGROUND: The core regulation of the abscisic acid (ABA) signalling pathway comprises the multigenic families PYL, PP2C, and SnRK2. In this work, we conducted a genome-wide study of the components of these families in Cucurbita pepo. RESULTS: The bioinformatic analysis of the C. pepo genome resulted in the identification of 19 CpPYL, 102 CpPP2C and 10 CpSnRK2 genes. The investigation of gene structure and protein motifs allowed to define 4 PYL, 13 PP2C and 3 SnRK2 subfamilies. RNA-seq analysis was used to determine the expression of these gene families in different plant organs, as well as to detect their differential gene expression during germination, and in response to ABA and cold stress in leaves. The specific tissue expression of some gene members indicated the relevant role of some ABA signalling genes in plant development. Moreover, their differential expression under ABA treatment or cold stress revealed those ABA signalling genes that responded to ABA, and those that were up- or down-regulated in response to cold stress. A reduced number of genes responded to both treatments. Specific PYL-PP2C-SnRK2 genes that had potential roles in germination were also detected, including those regulated early during the imbibition phase, those regulated later during the embryo extension and radicle emergence phase, and those induced or repressed during the whole germination process. CONCLUSIONS: The outcomes of this research open new research lines for agriculture and for assessing gene function in future studies.

Development of an In-Field Real-Time LAMP Assay for Rapid Detection of Tomato Leaf Curl New Delhi Virus.

Tomato leaf curl New Delhi virus (ToLCNDV) represents a threat to economically important horticultural crops. A real-time loop-mediated isothermal amplification (LAMP) assay for in-field ToLCNDV detection was developed, coupled to a rapid sample preparation method, and tested both in field and laboratory conditions on zucchini squash, tomato, and pepper samples. A set of six LAMP primers was designed for specific ToCLNDV detection, targeting a 218-nucleotide sequence within the AV1 gene. The sensitivity, specificity and accuracy of the real-time LAMP assay and comparison with canonical PCR were evaluated. The real-time LAMP assay developed was about one-thousand times more sensitive than the conventional PCR method, detecting a total of 4.41 × 102 genome copies as minimum target; no cross-reactivity was detected with the other geminiviruses used as the outgroup. The rapid sample preparation method allows for a reliable detection with a low reaction delay (≈2-3 min) compared to canonical DNA extraction, providing results in less than 45 min. Lastly, an increase in ToLCNDV-positive sample detection was observed compared to PCR, in particular for asymptomatic plants (85% and 71.6%, respectively). The real-time LAMP assay developed is a rapid, simple, specific, and sensitive technique for ToLCNDV detection, and it can be adopted as a routine test, for both in-field and laboratory conditions.

Effects of Below-Ground Microbial Biostimulant Trichoderma harzianum on Diseases, Insect Community, and Plant Performance in Cucurbita pepo L. under Open Field Conditions.

Agrochemicals are generally used in agriculture to maximize yields and product quality, but their overuse can cause environmental pollution and human health problems. To reduce the off-farm input of chemicals, numerous biostimulant products based on beneficial symbiont plant fungi are receiving a great deal of attention. The evolution of plant diseases and the performance of insects are influenced by plant chemical defences, both of which are, in turn, influenced by below-ground symbionts. Direct and indirect plant defences mediated by belowground symbionts against plant diseases and insect herbivores were demonstrated in greenhouses experiments. However, little attention has been paid to the use of Trichoderma under open field conditions, and no data are available for zucchini (Cucurbita pepo L.) plants in the field. To determine the effects of a commercial Trichoderma harzianum strain T22 on plant viruses, powdery mildew, the arthropod community, and on the agronomic performance associated with zucchini plants, an experiment was conducted in 2022 under open field conditions in South Italy. Our results indicate that T. harzianum T22 makes zucchini plants more attractive to aphids and to Hymenoptera parasitoid but failed to control zucchini pathogens. The complex plant-disease-arthropod-microorganism interactions that occurred in the field during the entire plant cycle are discussed to enrich our current information on the possibilities of using these microorganisms as a green alternative in agriculture.

Rearing Frankliniella zucchini Nakahara & Monteiro (Thysanoptera: Thripidae) on zucchini (Cucurbita pepo L. 'Caserta') fruits

Abstract Frankliniella zucchini transmits zucchini lethal chlorosis virus, causal agent of lethal chlorosis of zucchini squash. The characteristics of relationship between this virus with its vector have not been studied, one of the reasons being the lack of a method for rearing the thrips for laboratory studies. This work proposes a system for the rearing of F. zucchini on fresh virus free zucchini 'Caserta' fruits, offering a practical and efficient alternative for the supply of a large number of insects for later study of virus/vector relationship. In addition, to aid in the identification of this species of thrips, the immature and adult forms obtained from the colony were described.

Evaluation of Bioinsecticides for Management of Bemisia tabaci (Hemiptera: Aleyrodidae) and the Effect on the Whitefly Predator Delphastus catalinae (Coleoptera: Coccinellidae) in Organic Squash.

Organic zucchini squash is a high-value vegetable crop in Florida and potential exists to expand its production throughout the state. A lack of knowledge on the effectiveness of organic products and their integration with natural enemies is an important constraint to the regulation of pest populations in organic squash production in Florida. The objectives of this study were to evaluate the effect of insecticides labeled for organic production that can be used for management of Bemisia tabaci (Gennadius) biotype B, on organically grown squash; and to determine the effects of the most efficient insecticides on a key natural enemy, Delphastus catalinae (Horn). Experiments were conducted in the greenhouse in exclusion cages. The first experiment compared the effects of four bioinsecticides on whitefly densities. Insecticides include 1) AzaSol (azadirachtin), 2) PyGanic EC 1.4 (pyrethrin), 3) M-Pede (insecticidal soap), and 4) Entrust (spinosad). The second experiment investigated the effects of bioinsecticides on D. catalinae Treatment effectiveness was evaluated 1, 3, and 5 d posttreatment. PyGanic and M-Pede were highly effective in controlling whitefly populations on organic squash, while moderate control was provided by AzaSol and there was no control provided by Entrust. PyGanic and M-Pede treatments reduced D. catalinae populations when adults were released 1 d post pesticide application. However, when adults were released 5 d post application, there was no reduction. The importance of using bioinsecticides in combination with natural enemies to regulate pest populations in organic cropping systems is discussed.

Preference of Bemisia tabaci biotype B on zucchini squash and buckwheat and the effect of Delphastus catalinae on whitefly populations.

BACKGROUND: Zucchini squash, Cucurbita pepo L., is an important vegetable crop in Florida. Physiological disorders and insect-transmitted diseases are major problems for squash growers in semi-tropical regions around the world. Bemisia tabaci (Gennadius) biotype B is a significant whitefly pest and is largely responsible for transmitting viruses and causing physiological disorders in squash. Several studies have shown that whitefly populations are reduced when crops are interplanted with non-host cover crops or mulches. The aim of the present study was to determine how the presence of buckwheat, Fagopyrum esculentum Moench, and a key predator, Delphastus catalinae (Horn), affect whitefly colonization on squash. RESULTS: Whitefly densities were higher on squash than on buckwheat. The introduction of D. catalinae on squash significantly reduced whitefly populations. Overall, there were higher densities of D. catalinae on squash where the whitefly pest was more concentrated compared with buckwheat. CONCLUSION: The study provided preliminary evidence that D. catalinae, when used in conjunction with buckwheat as a living mulch, may aid in reducing whiteflies in squash. This greenhouse experiment highlights the need to investigate a multitactic approach of intercropping buckwheat with squash and the incorporation of D. catalinae in the field to manage populations of whiteflies and whitefly-transmitted diseases. © 2015 Society of Chemical Industry.