Effects associated with insertion of rol genes on morphogenic potential in explants derived from transgenic Bacopa monnieri (L.) Wettst

Mapping Intimacies ◽

10.21203/rs.3.rs-161892/v1 ◽

2021 ◽

Author(s):

Sayantika Sarkar ◽

Sumita Jha

Keyword(s):

Transgenic Plants ◽

Basal Medium ◽

Bacopa Monnieri ◽

Root Tip ◽

Root Explants ◽

Transformed Plants ◽

Root Organogenesis ◽

Number Of Leaves ◽

First Time ◽

Important Medicinal Plant

Abstract The present study deals with the establishment of rolA-transgenic and rolB-transgenic plants for the first time through Agrobacterium tumefaciens mediated transformation, exploiting the inherent morphogenic potential of an important medicinal plant, Bacopa monnieri (L.) Wettst. The rolA-transgenic and rolB-transgenic plants showed integration and expression of rolA and rolB genes respectively, whereas Ri-transformed plants showed integration and expression of rolA, rolB, rolC and rolD genes. Morphogenic potential of different types of explants derived from rolA-transgenic, rolB-transgenic and Ri-transformed plants on basal medium was evaluated. Shoot organogenesis was enhanced significantly in leaf (1.6-fold) and internode (1.4-fold) explants derived from rolA-transgenic plants, rolB-transgenic leaf (2.4-fold) and internode (1.6-fold) explants as well as leaf (5.2-fold) and internode (3.3-fold) explants derived from Ri-transformed plants compared to explants from non-transformed plants. Substantial increase in root organogenesis was also noticed in rolA-transgenic leaf (1.7-fold) explants, rolB-transgenic leaf (3.6-fold) and internode (1.4-fold) explants as well as leaf (4.1-fold) and internode (1.9-fold) explants derived from Ri-transformed plants compared to non-transformed ones. In addition to this, growth of root tip and shoot regeneration was also noticed from Ri-transformed root explants, but not in rolA-transgenic, rolB-transgenic and non-transformed roots. Clones of all three transgenic plants differed morphologically from non-transformed plants; however, rolB gene alone has a pronounced effect in alteration of plant phenotype in B. monnieri. Clones of rolB-transgenic plants were similar in shoot length, internode length, number of nodes/plant and number of leaves/plant when compared with Ri-transformed plant clones

Download Full-text

Somatic Embryogenesis and In vitro Plant Regeneration in Vigna trilobata (L.) Verd. - A Potential Pasture Legume

Plant Tissue Culture and Biotechnology ◽

10.3329/ptcb.v19i1.4990 ◽

1970 ◽

Vol 19 (1) ◽

pp. 89-99

Author(s):

K. Choudhary ◽

M. Singh ◽

M. S. Rathore ◽

N. S. Shekhawat

Keyword(s):

Somatic Embryogenesis ◽

Growth Regulators ◽

Somatic Embryos ◽

Basal Medium ◽

Long Term Study ◽

Continuous Application ◽

First Time ◽

Pasture Legume

This long term study demonstrates for the first time that it is possible to propagate embryogenic Vigna trilobata and to subsequently initiate the differentiation of embryos into complete plantlets. Initiation of callus was possible on 2,4-D. Somatic embryos differentiated on modified MS basal nutrient medium with 1.0 mg/l of 2,4-D and 0.5 mg/l of Kn. Sustained cell division resulted in globular and heart shape stages of somatic embryos. Transfer of embryos on to a fresh modified MS basal medium with 0.5 mg/l of Kn and 0.5 mg/l of GA3 helped them to attain maturation and germination. However, the propagation of cells, as well as the differentiation of embryos, were inhibited by a continuous application of these growth regulators. For this reason, a long period on medium lacking these growth regulators was necessary before the differentiation of embryos occurred again. The consequences for improving the propagation of embryogenic cultures in Vigna species are discussed. Key words: Pasture legume, Vigna trilobata, Globular, Heart shape, somatic embryogenesis D.O.I. 10.3329/ptcb.v19i1.4990 Plant Tissue Cult. & Biotech. 19(1): 89-99, 2009 (June)

Download Full-text

Overexpression of DUF538 from Wild Arachis Enhances Plant Resistance to Meloidogyne spp.

Agronomy ◽

10.3390/agronomy11030559 ◽

2021 ◽

Vol 11 (3) ◽

pp. 559

Author(s):

Ana Claudia Guerra Araujo ◽

Patricia Messenberg Guimaraes ◽

Ana Paula Zotta Mota ◽

Larissa Arrais Guimaraes ◽

Bruna Medeiros Pereira ◽

...

Keyword(s):

Transgenic Plants ◽

Redox System ◽

Redox Balance ◽

Uncharacterized Protein ◽

Chlorophyll Breakdown ◽

Domain Of Unknown Function ◽

Expression Behavior ◽

Meloidogyne Spp ◽

Wild Peanut ◽

First Time

DUF538 proteins belong to a large group of uncharacterized protein families sharing the highly conserved Domain of Unknown Function (DUF). Attention has been given to DUF538 domain-containing proteins due to changes in their gene expression behavior and protein abundance during plant development and responses to stress. Putative roles attributed to DUF538 in plants under abiotic and biotic constraints include involvement in cell redox balance, chlorophyll breakdown and pectin degradation. Our previous transcriptome studies suggested that DUF538 is also involved in the resistance responses of wild Arachis species against the highly hazardous root-knot nematodes (RKNs). To clarify the role of the AsDUF538 gene from the wild peanut relative Arachis stenosperma in this interaction, we analyzed the effect of its overexpression on RKN infection in peanut and soybean hairy roots and Arabidopsis transgenic plants. AsDUF538 overexpression significantly reduced the infection in all three heterologous plant systems against their respective RKN counterparts. The distribution of AsDUF538 transcripts in RKN-infected Arachis roots and the effects of AsDUF538 overexpression on hormonal pathways and redox system in transgenic Arabidopsis were also evaluated. This is the first time that a DUF538 gene is functionally validated in transgenic plants and the earliest report on its role in plant defense against RKNs.

Download Full-text

Agrobacterium tumefaciens-mediated Transformation of Double Haploid Canola (Brassica napus) Lines

Functional Plant Biology ◽

10.1071/pp96024 ◽

1997 ◽

Vol 24 (1) ◽

pp. 97 ◽

Cited By ~ 4

Author(s):

K. Kazan ◽

M. D. Curtis ◽

K. C. Goulter ◽

J. M. Manners

Keyword(s):

Brassica Napus ◽

Agrobacterium Tumefaciens ◽

Transgenic Plants ◽

Genetic Transformation ◽

Double Haploid ◽

Gene Promoter ◽

Root Explants ◽

Hypocotyl Explants ◽

Peroxidase Gene ◽

Hypocotyl Segments

Double haploid (DH) genotypes of canola (Brassica napus L.) have a high level of genetic uniformity but have not been previously tested for genetic transformation. Transgenic plants from three of four DH genotypes derived from cv. Westar were obtained by inoculation of either hypocotyl segments or root explants with Agrobacterium tumefaciens. For hypocotyl transformation, A. tumefaciens strain LBA4404 containing a binary plasmid with the neomycin phosphotransferase gene (nptII) and a CaMV 35S-peroxidase gene cassette was co-cultivated with hypocotyl segments taken from the 5–6-day-old seedlings. Transformation frequencies for hypocotyl explants of two DH genotypes were 0.3–3%. Direct evidence for genetic transformation of hypocotyl explants was obtained through molecular hybridisation analysis. Using this protocol, mature transformed plants were obtained within 4–6 months of co-cultivation. A method of root transformation was successfully modified for one DH genotype of canola and transgenic plants were obtained at a frequency of 2%. Using this protocol, a peroxidase gene promoter–GUS fusion construct was introduced into a DH genotype. Tissue specific GUS expression driven by the peroxidase gene promoter in transgenic plants was analysed by GUS staining. Transformation systems for double haploid canola lines will permit the assessment of introduced genes for their effect on agronomic and physiological traits.

Download Full-text

Mass Propagation of Nauclea diderrichii (de Willd. & Th. Dur) Merr. Seedlings through Tissue Culture Technique

Plant Tissue Culture and Biotechnology ◽

10.3329/ptcb.v31i1.54111 ◽

2021 ◽

Vol 31 (1) ◽

pp. 51-60

Author(s):

RI Oyediran ◽

JO Afolabi ◽

DB Olomola ◽

FO Akanni

Keyword(s):

Tissue Culture ◽

Adventitious Shoots ◽

Basal Medium ◽

Culture Technique ◽

Seedling Production ◽

Mass Propagation ◽

In Vitro Plantlets ◽

Number Of Leaves ◽

Nauclea Diderrichii

Nauclea diderrichii is a tree species of economic importance. However, its plantation establishment is limited by inadequate seedling production. Hence, there is ample scope of tissue culture for its mass propagation. Its in vitro plantlets development as affected by media strengths indicated that 100 % seed germination was obtained in full MS basal medium while the least (3.35 %) was from quarter-strength at 8 Weeks after inoculation (WAI). The effects of BAP and NAA assessed on the growth of its sub-cultured plantlets showed that highest number of leaves (17) and adventitious shoots (3) were obtained from MS basal medium supplemented with 0.1 mg/l BAP only. Whereas, highest shoot length (3.61 cm) and average number of roots (5/plantlet) were obtained from the same medium without hormone(s) at 8 WAI. Further sub-culturing into MS with 0.05 mg/l NAA resulted into plantlets having optimum shoot and massive root growth ready for acclimatization in 6 WAI. The plantlets were successfully acclimatized using coconuthusk/ topsoil mixture with 90 % survival. Plant Tissue Cult. & Biotech. 31(1): 51-60, 2021 (June)

Download Full-text

Unusual Mitochondrial Aggregation with Virus in Infected Transgenic Plants

Microscopy and Microanalysis ◽

10.1017/s1431927600026015 ◽

1998 ◽

Vol 4 (S2) ◽

pp. 1178-1179

Author(s):

L. Zhang ◽

W. G. Langenberg

Keyword(s):

Transgenic Plants ◽

Transgenic Plant ◽

Potato Virus Y ◽

Plant Cells ◽

Rna Sequences ◽

Transformed Plants ◽

Field Release ◽

The Family ◽

Positive Sense ◽

Ultrastructural Appearance

Viruses grouped in the family Potyviridaeare long flexious rod-shaped viruses and cause the most damage of all plant vims groups. They possess a single stranded positive sense RNA genome that encodes a single polyprotein product which yields eight or more proteins by secondary processing. Considerable efforts have been made to transform plants with DNA or RNA sequences of these proteins in hopes of obtaining pathogen-derived resistant plants. Hull has suggested that a close assessment of the risks involved in field release of transformed plants would be desirable. However, to our knowledge, there have been no published reports regarding properties of viruses that do infect some transformed plants. We describe here an ultrastructural appearance of transgenic plant cells infected by potato virus Y (PVY) (ATCC PV No. 50; PVY-50) and unusual mitochondrial aggregation with the virus.

Download Full-text

Root Colonization by Phenazine-1-Carboxamide-Producing Bacterium Pseudomonas chlororaphis PCL1391 Is Essential for Biocontrol of Tomato Foot and Root Rot

Molecular Plant-Microbe Interactions ◽

10.1094/mpmi.2000.13.12.1340 ◽

2000 ◽

Vol 13 (12) ◽

pp. 1340-1345 ◽

Cited By ~ 220

Author(s):

Thomas F. C. Chin-A-Woeng ◽

Guido V. Bloemberg ◽

Ine H. M. Mulders ◽

Linda C. Dekkers ◽

Ben J. J. Lugtenberg

Keyword(s):

Root Rot ◽

Root Colonization ◽

Root Tip ◽

Pseudomonas Chlororaphis ◽

Wild Type ◽

Animal Tissues ◽

Tomato Seedling ◽

Antifungal Metabolites ◽

Seedling Inoculation ◽

First Time

The phenazine-1-carboxamide-producing bacterium Pseudomonas chlororaphis PCL1391 controls tomato foot and root rot caused by Fusarium oxysporum f. sp. radicis-lycopersici. To test whether root colonization is required for biocontrol, mutants impaired in the known colonization traits motility, prototrophy for amino acids, or production of the site-specific recombinase, Sss/XerC were tested for their root tip colonization and biocontrol abilities. Upon tomato seedling inoculation, colonization mutants of strain PCL1391 were impaired in root tip colonization in a gnotobiotic sand system and in potting soil. In addition, all mutants were impaired in their ability to control tomato foot and root rot, despite the fact that they produce wild-type levels of phenazine-1-carboxamide, the antifungal metabolite previously shown to be required for biocontrol. These results show, for what we believe to be the first time, that root colonization plays a crucial role in biocontrol, presumably by providing a delivery system for antifungal metabolites. The ability to colonize and produce phenazine-1-carboxamide is essential for control of F. oxysporum f. sp. radicis-lycopersici. Furthermore, there is a notable overlap of traits identified as being important for colonization of the rhizosphere and animal tissues.

Download Full-text

Kanamycin in Cereal Biotechnology: A Screening or a Selectable Marker?

BioScientific Review ◽

10.32350/bsr.0301.01.i ◽

2021 ◽

Vol 3 (1) ◽

pp. 01-05

Author(s):

Malik Shuja

Keyword(s):

Transgenic Plants ◽

Genetic Transformation ◽

Selectable Marker ◽

Normal Growth ◽

Minimal Effect ◽

Effective Screening ◽

Transformed Plants ◽

Transformation Systems ◽

Selection Agent ◽

Screening Marker

Kanamycin is a widely used selection agent in dicot-plant genetic transformation systems. In monocots, however, it does not seem to be effective as it has no or minimal effect on the normal growth of non-transformed plants. Kanamycin was previously demonstrated to bleach the pigments of the non-transgenic plants. This may yield the idea that kanamycin can be used as an effective screening marker rather than a selectable marker in monocots.

Download Full-text

TDZ Plays Key Role in Shoot Regeneration from Different Explants of Picrorhiza kurroa: An Endangered Medicinal Herb of Western Himalayas

Current Journal of Applied Science and Technology ◽

10.9734/cjast/2019/v33i430089 ◽

2019 ◽

pp. 1-9 ◽

Cited By ~ 1

Author(s):

Vanita Patial ◽

Amita Bhattacharya

Keyword(s):

Natural Habitat ◽

Basal Medium ◽

In Vitro Rooting ◽

Western Himalayas ◽

Picrorhiza Kurroa ◽

Regeneration Potential ◽

Root Explants ◽

Shoot Number ◽

Endangered Medicinal Herb

Picrorhiza kurroa plants were collected from its natural habitat. In vitro plants were raised from the leaves of high yielding collection screened in an earlier study. Leaves, roots and internodal segments were cultured for 15 days. The effect of thidiazuron (1-phenyl-3-(1, 2, 3- thiadiazol-5-yl) urea; TDZ) pretreatment for 15 days on regeneration potential of different explants viz. leaves, roots and internodes of Picrorhiza kurroa was studied. Regeneration potential varied significantly with the type of explant. Regeneration response of 100% with 46.25 shoots per explant was obtained in leaf segments of 2.0 cm length pretreated with 0.5 µM TDZ for 15 days and then transferred to 2.32 µM kinetin (Kn) containing MS basal medium. In case of root explants maximum shoot number (17.12) was obtained on 0.5 µM TDZ pretreated for 15 days and then to 3.64 µM Kn. Maximum shoots per explants (12.33) were obtained in case of internodes pretreated with 0.5 µM TDZ for 15 days and transferred to 1.16 µM Kn. Regenerated shoots from different explants developed in vitro rooting on MS basal medium within 7-8 days. Conclusively, an efficient and repeatable protocol for rapid regeneration from different explants and in vitro rooting has been developed in P. kurroa which can be effectively used for its conservation.

Download Full-text

Evaluation of Resistance to Gummy Stem Blight (Didymella bryoniae) Disease in ipt (Isopentenyl Transferase) Transformed Watermelon Plants

HortScience ◽

10.21273/hortsci.39.4.820e ◽

2004 ◽

Vol 39 (4) ◽

pp. 820E-821

Author(s):

Fahrettin Goktepe ◽

Harrison Hughes*

Keyword(s):

Transgenic Plants ◽

Infected Leaf ◽

Wild Type ◽

Didymella Bryoniae ◽

Isopentenyl Transferase ◽

Disease Symptoms ◽

Stem Blight ◽

Transformed Plants ◽

Gummy Stem Blight ◽

Run Off

Watermelon plants are susceptible to Gummy stem blight disease that considerably reduces yields worldwide. In order to develop non-specific resistance, watermelon cv. Crimson Sweet was transformed with copper inducible isopentenyl transferase (ipt), the rate-limiting step in cytokinin biosynthesis, gene via Agrobacterium tumafaciences (LBA4404). Transformed (ipt) and nontransformed plants were grown at approximately 28-30 °C day, 20-22 °C night and 16 hours daylight under greenhouse conditions. Once the plants initiated new growth both transgenic plants and wild type plants were sprayed with one of three different concentrations (0, 10 & 50 μm) of CuSO4. Plants were sprayed twice to run-off in a twenty-four hour time period before inoculation with the pathogen. Cultures of the pathogen Didymela bryonia (W353) were grown for about 3 weeks and an inoculum containing 105 conidia per mL was sprayed with the prepared suspension until initial run-off in a humidified chamber. The disease symptoms were evaluated after one week with resistance demonstrated in all treated transgenic plants. All nonsprayed transgenic and wild type plants showed similar disease symptoms. Infected leaf samples were surface sterilized and re-cultured on V8 medium. The characteristics of the recovered pathogen confirmed that it was identical to the stock culture of W353. The same experiment has been conducted on seedlings from transgenic (T1 generations) and non-transformed plants. The non-transformed seedlings showed the first disease symptoms on their cotyledons and lower leaves. Disease resistance was observed in seedlings of the treated transformed plants as compared to nontransformed ones.

Download Full-text