scholarly journals Establishment of a Novel and Efficient Agrobacterium-Mediated in Planta Transformation System for Passion Fruit (Passiflora edulis)

Plants ◽  
2021 ◽  
Vol 10 (11) ◽  
pp. 2459
Author(s):  
Hafiz Muhammad Rizwan ◽  
Qiang Yang ◽  
Ahmed Fathy Yousef ◽  
Xiaoxue Zhang ◽  
Yasir Sharif ◽  
...  
Keyword(s):  
Virulent Strain ◽  
Crop Improvement ◽  
Multiple Shoots ◽  
Passion Fruit ◽  
In Planta Transformation ◽  
Transformation System ◽  
In Planta ◽  
Labor Costs ◽  
Passiflora Edulis ◽  
Functional Studies

Passion fruit (Passiflora edulis) is an important fruit crop with high economic value. Genetic engineering plays an important role in crop improvement with desired traits and gene functional studies. The lack of a simple, efficient, and stable transformation system for passion fruit has greatly limited gene functional studies. In this study, a simple and efficient Agrobacterium-mediated in planta transformation system for passion fruit was established, using Agrobacterium virulent strain EHA105 harboring the binary vectors pCAMBIA1301 and pCAMBIA1302 with GUS and GFP reporter genes. The system requires less time and labor costs than conventional transformation systems, and no additional phytohormones and sterile conditions are required. Regeneration efficiency of 86% and transformation efficiency of 29% were achieved, when the wounds were wrapped with Parafilm and the plants were kept in darkness for 15 days. Approximately 75% of the regenerated plants had a single shoot and 26% multiple shoots. The transformation was confirmed at the DNA and RNA levels as well as by GUS staining and GFP fluorescent measurements. The developed protocol will contribute to the genetic improvement of passion fruit breeding.

scholarly journals A simple and fast Agrobacterium-mediated transformation system for passion fruit KPF4 (Passiflora edulis f. edulis × Passiflora edulis f. flavicarpa)

Plant Methods ◽  
2020 ◽  
Vol 16 (1) ◽  
Author(s):  
Lydia K. Asande ◽  
Richard O. Omwoyo ◽  
Richard O. Oduor ◽  
Evans N. Nyaboga
Keyword(s):  
Transgenic Plants ◽  
Transformation Efficiency ◽  
Cell Transformation ◽  
Raw Materials ◽  
Leaf Disc ◽  
Fruit Production ◽  
Histochemical Staining ◽  
Passion Fruit ◽  
Transformation System ◽  
Passiflora Edulis

Abstract Background Passion fruit (Passiflora edulis Sims) is an important horticultural crop in the tropics and subtropics, where it has great commercial potential due to high demand for fresh edible fruits and processed juice as well as source of raw materials in cosmetic industries. Genetic engineering shows great potential in passion fruit improvement and can compensate for the limitations of conventional breeding. Despite the success achieved in genetic modification of few passion fruit varieties, transgenic passion fruit production is still difficult for farmer-preferred cultivars. Therefore, it is important to establish a simple and fast Agrobacterium-mediated cell transformation of commercial hybrid passion fruit KPF4 (Passiflora edulis f. edulis × Passiflora edulis f. flavicarpa). Results In the present study, we have developed a simple and fast Agrobacterium-mediated transformation system for hybrid passion fruit KPF4 using leaf disc explants. Factors affecting the rate of transient beta (β)-glucuronidase (gusA) expression and consequently transformation efficiency were optimized as follows: Agrobacterium cell density with an OD600 of 0.5, 30 min infection time, 3 days of co-cultivation duration and the incorporation of 200 µM acetosyringone into Agrobacterium infection suspension medium. Using the optimized conditions, transgenic plants of KPF4 were produced within 2 months with an average transformation efficiency of 0.67%. The β-glucuronidase (GUS) histochemical staining confirmed the expression and integration of an intron-containing gusA gene into transformed leaf discs and transgenic plant lines of KPF4. The presence of gusA gene in the transgenic plants was confirmed by polymerase chain reaction (PCR). The results confirmed that the gusA gene was efficiently integrated into the passion fruit genome. Conclusions The developed transformation protocol is simple and rapid and could be useful for functional genomic studies and transferring agronomically important traits into passion fruit hybrid KPF4. This study developed a method that can be used to transfer traits such as resistance to viral diseases, low fruit quality and short storage life. To the best of our knowledge, this is the first report on genetic transformation system for commercial passion fruit hybrid KPF4.

Optimization and establishment of genotype-independent seed-based in planta transformation system in cotton (Gossypium hirsutum L.cv.SVPR-2)

Plant Gene ◽  
2021 ◽  
pp. 100296
Author(s):  
Prem Kumar Ganesan ◽  
Sivakumar Subiramani ◽  
Siva Govindarajan ◽  
Vigneswaran Manickam ◽  
Senthil Kumar Thiruppathi ◽  
...  
Keyword(s):  
Gossypium Hirsutum ◽  
In Planta Transformation ◽  
Transformation System ◽  
In Planta

scholarly journals A simple and fast Agrobacterium-mediated transformation system for hybrid passion fruit (Passiflora edulis f. edulis × Passiflora edulis f. flavicarpa)

2020 ◽  
Author(s):  
Lydia K. Asande ◽  
Richard O. Omwoyo ◽  
Richard O. Oduor ◽  
Evans Nyaboga
Keyword(s):  
Passion Fruit ◽  
Transformation System ◽  
Passiflora Edulis ◽  
Agrobacterium Mediated Transformation

Abstract The authors have removed this preprint from Research Square.

In planta transformation: A smart way of crop improvement

2020 ◽  
pp. 351-362 ◽  
Author(s):  
Shabnam K. Saifi ◽  
Nishat Passricha ◽  
Renu Tuteja ◽  
Pushpa Kharb ◽  
Narendra Tuteja
Keyword(s):  
Crop Improvement ◽  
In Planta Transformation ◽  
In Planta

scholarly journals In Planta Transformation in Plants: A Review

2019 ◽  
Vol 40 (03) ◽  
Author(s):  
Ratna Preeti Kaur ◽  
Sugani Devi
Keyword(s):  
Tissue Culture ◽  
Developmental Stages ◽  
Crop Improvement ◽  
Induction Medium ◽  
Foreign Gene ◽  
In Planta Transformation ◽  
In Planta ◽  
Agrobacterium Strain ◽  
Plant Parts ◽  
Whole Plants

In planta transformation has been established as an innovative and simple technique involving direct transformation of plant parts without involving the tedious tissue culture step. Methodologies of in planta transformation involve different plant parts and strategies. Agrobacterium strain carrying the gene of interest is targeted to the specific plant part either directly or in the induction medium. Vacuum infiltration is sometimes used to facilitate foreign gene integration. Initial studies of this novel technique involved treatment of whole plants with the inoculum, and later shifted to treatments of shoot tips and floral parts and finally the female reproductive parts have been targeted. Zygotes, embryos and seeds have recently been used extensively yielding successful transformation. The method is simple, convenient and overcomes the problem of tissue culture induced genetic variability in the transformants. The review traces the origin and development in the in planta methodologies used over the past and the various parameters considered by the workers for increased effectiveness viz. developmental stages, Agrobacterium stain, surfactant, induction medium etc., in the various crops compiled. Based on the review it may be inferred that there is immense potential in planta transformation, and the ease of regeneration and selection of transformants in the methods described, can be utilized for crop improvement.

scholarly journals Biological fixation, transfer and balance of nitrogen in passion fruit (Passiflora edulis Sims) orchard intercropped with different green manure crops

2019 ◽  
Vol 13 ((03) 2019) ◽  
pp. 465-471 ◽  
Author(s):  
Edilândia Farias Dantas ◽  
Ana Dolores Santiago de Freitas ◽  
Maria do Carmo Catanho Pereira de Lyra ◽  
Carolina Etienne de Rosália e Silva Santos ◽  
Stella Jorge de Carvalho Neta ◽  
...  
Keyword(s):  
Cover Crops ◽  
Pigeon Pea ◽  
Passion Fruit ◽  
Biological Fixation ◽  
Mineral Fertilization ◽  
Passiflora Edulis ◽  
Jack Bean ◽  
Positive Balance ◽  
Sunn Hemp ◽  
Passiflora Edulis Sims

Green manures can replace or supplement mineral fertilization and add organic matter to the soils, ensuring greater sustainability to fruit growing in semiarid regions. Biological fixation, transfer and balance of nitrogen were determined on an irrigated yellow passion fruit orchard (Passiflora edulis Sims) intercropped separately with three cover crops: sunn hemp, Crotalaria juncea (L.); pigeon pea, Cajanus cajan (L.) Mill; and jack bean, Canavalia ensiformis (L.) DC. In a fourth treatment, legumes were not planted, but spontaneous vegetation was left to grow freely. The legumes were croped for 90 days in three lines (0.5 m apart) inside the passion fruit plant lines (2.5 m apart). Fixation and transfers were determined by the 15N natural abundance technique, using sunflower as a reference plant. The three planted legumes nodulated abundantly and fixed nitrogen in high proportions (between 50 and 90% of their N), forming symbiosis with bacteria naturally established in the soil. Jack bean produced more biomass than sunn hemp and pigeon pea, and as much as the spontaneous plants, of which 23% were legumes. The amounts of fixed N (150, 43, 30 and 29 kg ha-1) were determined mainly by the biomass of legumes. More than 40% of the N of passion fruit plants came from the biological nitrogen fixation of the intercropped jack bean, which provided an amount of N higher than that exported in the fruits, generating a positive balance of more than 100 kg ha-1. Therefore, it is recommended to intercrop jack bean in irrigated passion fruit orchards.

scholarly journals Genetic Diversity of Purple Passion Fruit, Passiflora edulis f. edulis, Based on Single-Nucleotide Polymorphism Markers Discovered through Genotyping by Sequencing

Diversity ◽  
2021 ◽  
Vol 13 (4) ◽  
pp. 144
Author(s):  
Nohra Castillo Rodríguez ◽  
Xingbo Wu ◽  
María Isabel Chacón ◽  
Luz Marina Melgarejo ◽  
Matthew Wohlgemuth Blair
Keyword(s):  
Genetic Diversity ◽  
Single Nucleotide Polymorphism ◽  
Genotyping By Sequencing ◽  
Passion Fruit ◽  
Nucleotide Polymorphism ◽  
Single Nucleotide ◽  
Passiflora Edulis ◽  
Tropical Fruit ◽  
Commercial Cultivars ◽  
Purple Passion Fruit

Orphan crops, which include many of the tropical fruit species used in the juice industry, lack genomic resources and breeding efforts. Typical of this dilemma is the lack of commercial cultivars of purple passion fruit, Passiflora edulis f. edulis, and of information on the genetic resources of its substantial semiwild gene pool. In this study, we develop single-nucleotide polymorphism (SNP) markers for the species and show that the genetic diversity of this fruit crop has been reduced because of selection for cultivated genotypes compared to the semiwild landraces in its center of diversity. A specific objective of the present study was to determine the genetic diversity of cultivars, genebank accession, and landraces through genotyping by sequencing (GBS) and to conduct molecular evaluation of a broad collection for the species P. edulis from a source country, Colombia. We included control genotypes of yellow passion fruit, P. edulis f. flavicarpa. The goal was to evaluate differences between fruit types and compare landraces and genebank accessions from in situ accessions collected from farmers. In total, 3820 SNPs were identified as informative for this diversity study. However, the majority distinguished yellow and purple passion fruit, with 966 SNPs useful in purple passion fruits alone. In the population structure analysis, purple passion fruits were very distinct from the yellow ones. The results for purple passion fruits alone showed reduced diversity for the commercial cultivars while highlighting the higher diversity found among landraces from wild or semi-wild conditions. These landraces had higher heterozygosity, polymorphism, and overall genetic diversity. The implications for genetics and breeding as well as evolution and ecology of purple passion fruits based on the extant landrace diversity are discussed with consideration of manual or pollinator-assisted hybridization of this species.

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