scholarly journals Regeneration and Genetic Transformation of Arabidopsis Pumila: An Ephemeral Plant Suitable for Investigating the Mechanisms for Adaptation to Desert Environments

2021 ◽  
Author(s):  
Yuhuan Jin ◽  
Li Guo ◽  
Danqing Liu ◽  
Yongguang Li ◽  
Hao Ai ◽  
...  
Keyword(s):  
Adventitious Shoots ◽  
Cellular Level ◽  
Cauliflower Mosaic Virus ◽  
Induction Medium ◽  
35S Promoter ◽  
Naphthalene Acetic Acid ◽  
Growth Duration ◽  
Petiole Explants ◽  
Induction Rate ◽  
Ephemeral Plant

Abstract Arabidopsis pumila is a type of cruciferous ephemeral plant, which in China mainly grows in the desert environments of northern Xinjiang. A. pumila not only has a short growth duration, but also has high photosynthetic efficiency, seed yield, salt tolerance, and drought resistance. It is an ideal species for the study of environmental adaptations in ephemeral plants. We induced callus tissue formation on the roots and hypocotyls of 8-day-old seedlings, and on the leaves and petioles of 4-week-old seedlings, and obtained multiple adventitious shoots on these tissues grown on Murashige and Skoog induction medium supplemented with 0.5 mg/L 6-Benzylaminopurine and 0.1 mg/L α-Naphthalene acetic acid. Young roots, hypocotyls, leaves, and petioles could all induce calluses, but the induction rate was highest on young roots. In addition, the leaves and petioles of 4-week-old seedlings were used as explants, the Δ1-pyrroline-5-carboxylic acid synthase gene 1 of A. pumila controlled by 35S promoter of cauliflower mosaic virus was used as target gene, and hygromycin B was used as screening antibiotic to explore Agrobacterium tumefaciens GV3101 mediated transformation. The results showed that the callus induction rate of petiole explants was the highest when they were treated with Agrobacterium suspension (OD600 = 0.6) for 10 min and thenco-cultured in dark for 2 d. The qRT-PCR results showed that the ApP5CS1.1 gene was overexpressed in the transgenic plants. These protocols provide working research methods for exploring the cellular level adaptative mechanisms of this species to desert environments.

scholarly journals Plant Regeneration through Protocorm-like Bodies Induced from Nodal Explants of Syngonium podophyllum ‘White Butterfly’

HortScience ◽  
2008 ◽  
Vol 43 (7) ◽  
pp. 2129-2133 ◽  
Author(s):  
Jin Cui ◽  
Juanxu Liu ◽  
Min Deng ◽  
Jianjun Chen ◽  
Richard J. Henny
Keyword(s):  
Adventitious Shoots ◽  
Leaf Explants ◽  
Basal Medium ◽  
Shoot Culture ◽  
Nodal Explants ◽  
Induction Medium ◽  
Naphthalene Acetic Acid ◽  
Ex Vitro ◽  
Petiole Explants

Syngonium podophyllum ‘White Butterfly’, one of the most popular ornamental foliage plants, is propagated almost exclusively through in vitro shoot culture. Ex vitro rooting, however, has been associated with severe Myrothecium leaf spot (Myrothecium roridum Tode ex Fr.). The objective of this study was to establish a method for regenerating well-rooted plantlets before ex vitro transplanting. Leaf and petiole explants were cultured on a Murashige and Skoog (MS) basal medium supplemented with N-(2-chloro-4-pyridyl)-N′-phenylurea (CPPU), N-phenyl-N′-1,2,3-thiadiazol-5-ylurea (TDZ), 6-benzyladenine (BA), or N-isopentenylaminopurine (2iP) with α-naphthalene acetic acid (NAA) and 2,4-dichlorophenoxyacetic acid (2,4-D), respectively. Calli formed from leaf explants cultured on the basal medium supplemented with CPPU or TDZ with 2,4-D or with NAA as well as from petiole explants cultured on the medium supplemented with BA, CPPU, or TDZ with 2,4-D or NAA. The calli, however, failed to differentiate, and shoot organogenesis did not occur. Culture of nodal explants on the MS basal medium supplemented with 9.84 μm 2iP, 8.88 μm BA, 8.07 μm CPPU, or 9.08 μm TDZ with 2.26 μm 2,4-D resulted in the formation of protocorm-like bodies, adventitious shoots, and subsequently well-rooted plantlets. MS basal medium supplemented with 19.68 μm 2iP and 1.07 μm NAA resulted in the highest percentage (92.9%) of nodal explants producing protocorm-like bodies and an average of 16.9 well-rooted plantlets per nodal explant. Adventitious shoots were able to root in the initial induction medium, but better root development occurred after shoots with protocorm-like bodies were transferred onto MS basal medium supplemented with 9.84 μm 2iP and 2.69 μm NAA. Regenerated plantlets were stable and grew vigorously with 100% survival rates after ex vitro transplanting to a container substrate in a shaded greenhouse.

Surface structural analysis of cultured cotyledons of Douglas-fir

1982 ◽  
Vol 60 (12) ◽  
pp. 2729-2733 ◽  
Author(s):  
Edward G. Kirby ◽  
Margaretha E. Schalk
Keyword(s):  
Adventitious Shoots ◽  
Adventitious Shoot ◽  
Shoot Formation ◽  
Douglas Fir ◽  
Induction Medium ◽  
Naphthalene Acetic Acid ◽  
Shoot Induction ◽  
Adventitious Shoot Formation ◽  
Callus Proliferation ◽  
Shoot Induction Medium

Cotyledons of Douglas-fir are triangular in cross section and possess two epistomatic surfaces with centrally located stomatal rows (commonly seven). After 1 week in culture on a medium inducing adventitious shoot formation (5 μM N6-benzylaminopurine and 5 μM α-naphthalene acetic acid (NAA)) or callus proliferation (5 μM NAA) cells of the hypodermal region immediately below the epidermis begin to elongate and divide rupturing the epidermis. Apical domes of adventitiously produced bud primordia emerge from the ruptured epidermis after 14–21 days in culture on shoot induction medium. Emergence of buds takes place preferentially on epistomatic cotyledonary surfaces. The large number of hypodermal cells that respond to shoot induction medium by forming adventitious shoots suggests further investigation of fundamental events associated with morphogenesis in cotyledon cultures of Douglas-fir.

Developing a scale-up system for the in vitro multiplication of thidiazuron-induced strawberry shoots using a bioreactor

2008 ◽  
Vol 88 (4) ◽  
pp. 737-746 ◽  
Author(s):  
Samir C Debnath
Keyword(s):  
Large Scale ◽  
Scale Up ◽  
Adventitious Shoots ◽  
Shoot Proliferation ◽  
Shoot Multiplication ◽  
Induction Medium ◽  
Fragaria Ananassa ◽  
Petiole Explants ◽  
Bioreactor System

The use of large-scale liquid cultures in a bioreactor system has the potential to resolve the manual handling of the various stages of micropropagation and increases shoot multiplication in vitro significantly compared with those cultured on semi-solid gelled medium. In an attempt to improve the micropropagation protocol for strawberry (Fragaria × ananassa Duch.), a procedure for the mass propagation of adventitious shoots regenerated from leaf, sepal and petiole explants of cultivar Bounty using a liquid medium-containing bioreactor system combined with gelled medium is described. Leaf disks, sepals and petiole halves produced multiple buds and shoots without an intermediary callus phase on 2-4 µM thidiazuron (TDZ)-containing shoot induction medium within 5-6 wk of culture initiation. TDZ supported rapid shoot proliferation at low concentrations (0.1 µM), but induced hyperhydricity in a bioreactor system. Bioreactor-multiplied hyperhydric shoots were transferred to gelled medium containing 2-4 µM zeatin, and produced normal shoots and root within 4 wk of culture. In vitro derived plantlets were acclimatized and eventually established in the greenhouse and in the field. Present results suggested the possibility of large-scale multiplication of strawberry shoots in bioreactors. Key words: Fragaria × ananassa, growth regulator, shoot regeneration, RITA® bioreactor

scholarly journals In Vitro Propagation, Huperzine A Content and Antioxidant Activity of Three Genotypic Huperzia serrata

Plants ◽  
2021 ◽  
Vol 10 (6) ◽  
pp. 1112
Author(s):  
Yan Yang ◽  
Liangfang Dai ◽  
Decai Wu ◽  
Limin Dong ◽  
Yisheng Tu ◽  
...  
Keyword(s):  
Antioxidant Activity ◽  
High Performance ◽  
In Vitro Propagation ◽  
Huperzine A ◽  
Induction Medium ◽  
Naphthalene Acetic Acid ◽  
Regeneration Rate ◽  
Huperzia Serrata ◽  
Chromatography Analysis

Huperzia serrata is a traditional herb and endangered Chinese medicinal material, which has attracted much attention due to its production of Huperzine A (HupA). In vitro propagation of H. serrata is considered a new way to relieve the resource pressure of H. serrata. In this study, three different genotypic wild H. serrata were used for in vitro propagation. Then, the antioxidant activity and the content of HupA in the regenerated H. serrata were investigated. The results showed the survival rate of the explant was increased to 25.37% when using multiple sterilization processes. The best induction medium for H. serrata was the Schenk and Hildebrandt (SH) medium supplemented with 0.5 mg·L−1 Naphthalene acetic acid (NAA) and 0.1 mg·L−1 2,4-Dichlorophenoxyacetic acid (2,4-D), where the regeneration rate of the explant was to 57.04%. The best proliferation medium was the SH medium with NAA (1.0 mg·L−1), as the biomass of in vitro tissue increased 164.17 ± 0.41 times. High-performance liquid chromatography analysis showed that the in vitro culture of three genotypes could produce HupA and the content of HupA was 53.90–87.17 µg·g−1. The antioxidant experiment showed that the methanol extract of in vitro H. serrata had higher antioxidant activity than that of wild H. serrata. This study provides a reliable in vitro H. serrata culture protocol and laid an important foundation for the antioxidant capacity of the thallus and the content of HupA.

scholarly journals Overexpression of the Golden SNP-Carrying Orange Gene Enhances Carotenoid Accumulation and Heat Stress Tolerance in Sweetpotato Plants

Antioxidants ◽  
2021 ◽  
Vol 10 (1) ◽  
pp. 51
Author(s):  
So-Eun Kim ◽  
Chan-Ju Lee ◽  
Sul-U Park ◽  
Ye-Hoon Lim ◽  
Woo Sung Park ◽  
...  
Keyword(s):  
Heat Stress ◽  
Stress Tolerance ◽  
Ipomoea Batatas ◽  
Radical Scavenging Activity ◽  
Radical Scavenging ◽  
Cauliflower Mosaic Virus ◽  
Carotenoid Content ◽  
35S Promoter ◽  
Storage Roots ◽  
Heat Stress Tolerance

Carotenoids function as photosynthetic accessory pigments, antioxidants, and vitamin A precursors. We recently showed that transgenic sweetpotato calli overexpressing the mutant sweetpotato (Ipomoea batatas [L.] Lam) Orange gene (IbOr-R96H), which carries a single nucleotide polymorphism responsible for Arg to His substitution at amino acid position 96, exhibited dramatically higher carotenoid content and abiotic stress tolerance than calli overexpressing the wild-type IbOr gene (IbOr-WT). In this study, we generated transgenic sweetpotato plants overexpressing IbOr-R96H under the control of the cauliflower mosaic virus (CaMV) 35S promoter via Agrobacterium-mediated transformation. The total carotenoid contents of IbOr-R96H storage roots (light-orange flesh) and IbOr-WT storage roots (light-yellow flesh) were 5.4–19.6 and 3.2-fold higher, respectively, than those of non-transgenic (NT) storage roots (white flesh). The β-carotene content of IbOr-R96H storage roots was up to 186.2-fold higher than that of NT storage roots. In addition, IbOr-R96H plants showed greater tolerance to heat stress (47 °C) than NT and IbOr-WT plants, possibly because of higher DPPH radical scavenging activity and ABA contents. These results indicate that IbOr-R96H is a promising strategy for developing new sweetpotato cultivars with improved carotenoid contents and heat stress tolerance.

Aberrant promoter methylation occurred from multicopy transgene and SU(VAR)3 - 9 homolog 9 (SUVH9) gene in transgenic Nicotiana benthamiana

2012 ◽  
Vol 39 (9) ◽  
pp. 764 ◽  
Author(s):  
Gi-Ho Lee ◽  
Seong-Han Sohn ◽  
Eun-Young Park ◽  
Young-Doo Park
Keyword(s):  
Gene Silencing ◽  
Nicotiana Benthamiana ◽  
Fluorescent Protein ◽  
Cauliflower Mosaic Virus ◽  
Camv 35S Promoter ◽  
35S Promoter ◽  
Camv 35S ◽  
Histone Deacetylase 1 ◽  
Transgenic Lines ◽  
Green Fluorescent

The chemical modification of DNA by methylation is a heritable trait and can be subsequently reversed without altering the original DNA sequence. Methylation can reduce or silence gene expression and is a component of a host’s defence response to foreign nucleic acids. In our study, we employed a plant transformation strategy using Nicotiana benthamiana Domin to study the heritable stability of the introduced transgenes. Through the introduction of the cauliflower mosaic virus (CaMV) 35S promoter and the green fluorescent protein (GFP) reporter gene, we demonstrated that this introduced promoter often triggers a homology-dependent gene-silencing (HDGS) response. These spontaneous transgene-silencing phenomena are due to methylation of the CaMV 35S promoter CAAT box during transgenic plant growth. This process is catalysed by SU(VAR)3–9 homologue 9 (SUVH9), histone deacetylase 1 (HDA1) and domains rearranged methylase 2 (DRM2). In particular, we showed from our data that SUVH9 is the key regulator of methylation activity in epigenetically silenced GFP transgenic lines; therefore, our findings demonstrate that an introduced viral promoter and transgene can be subject to a homology-dependent gene-silencing mechanism that can downregulate its expression and negatively influence the heritable stability of the transgene.

scholarly journals Ação de extratos vegetais e fitoreguladores na organogênese de Begonia rex

1998 ◽  
Vol 20 (20) ◽  
pp. 131
Author(s):  
Elcí Terezinha Henz Franco ◽  
Cinara Echart Almeida
Keyword(s):  
Acetic Acid ◽  
Plant Regeneration ◽  
Basal Medium ◽  
Optimal Combination ◽  
Coconut Water ◽  
Naphthalene Acetic Acid ◽  
Petiole Explants ◽  
Potato Extract ◽  
Whole Plants

Petiole explants of Begonia rex were cultured on basal medium (MURASHIGE & SKOOG, 1962). The medium was suplemented with naphthalene acetic acid (0.01; 0.1 and 0.5 mg/I) and kinetin ( 0.1; 0.2; 0.5 and 1.0 mg/I). In these experiments, were as also used coconut water (15% and 20%) or potato extract ( 15% and 20%). Buds were formed in several treatments, but the best combination was coconut water 0.01 mg/L NAA and 0.1 mg/I KIN. Whole plants (40% of the explants) were obtained when was added coconut water . The optimal combination for plant regeneration (100%) was 0.01 mg/I NAA plus 0.1 mg/I KIN.

scholarly journals In-vitro development of Nauclea diderrichii (de Willd. & Th. Dur) Merrin liquid-M Smedia supplemented with benzyl amino purine (BAP) and naphthalene acetic acid (NAA)

2021 ◽  
Vol 24 (12) ◽  
pp. 2065-2069
Author(s):  
J.O. Afolabi ◽  
R.I. Oyediran ◽  
Y.O. Aguda ◽  
E.A. Adekunle
Keyword(s):  
Growth Regulators ◽  
Adventitious Shoots ◽  
Transition Process ◽  
Multiple Shoot ◽  
Naphthalene Acetic Acid ◽  
Temporary Immersion Bioreactor ◽  
Group A ◽  
Bioreactor System ◽  
Vitro Development

The growth of plantlets in Temporary Immersion Bioreactor system (TIBs) relies on initial successful liquid phase transition process. The response of N. diderrichii explants was assessed in liquid-M Smedia with a view to mass produce its seedlings using TIBs. Seven treatments consisting (A) 0.0/0.0, (B) 0.0/0.1, (C) 0.1/0.0, (D) 0.2/0.1, (E) 0.3/0.2, (F) 0.4/0.3 and (G) 0.5/0.4mg/lBAP/NAA combinations were studied. Each group consist of seven replicates and group A without Growth Regulators (GR) serves as control. The results at 4 Weeks after Inoculation (WAI) showed that effects of the growth regulators were significant on shoot length and number of adventitious shoots while number of roots and leaves were closely related. Treatment E produced highest number of adventitious shoots (3.6) which was higher than 0.9 shoots from treatment G and closely related to others. Maximum number of leaves (16.6) was produced by treatment F followed by E (15.7) while the least (12) was obtained in treatment A. The highest number of roots (4.9) was obtained from treatments B, followed by E (4.3) with the lowest being recorded in C (2.43). Liquid MS medium supplemented with 0.3/0.2mg/lBAP/NAA shows some promise for plantlets generation for the purpose of multiple shoot production of N. diderrichii in TIBs.

Chloroplast-dependent and light-independent expression of the tobacco rbcS promoter–GUS chimeric gene in black spruce

1996 ◽  
Vol 26 (6) ◽  
pp. 909-917
Author(s):  
Madoka Gray-Mitsumune ◽  
Bong Y. Yoo ◽  
Pierre J. Charest
Keyword(s):  
Fusion Gene ◽  
Black Spruce ◽  
Gus Activity ◽  
Chimeric Gene ◽  
Cauliflower Mosaic Virus ◽  
35S Promoter ◽  
Zygotic Embryos ◽  
Ribulose Bisphosphate Carboxylase ◽  
Bisphosphate Carboxylase ◽  
Rbcs Promoter

The tobacco rbcS (ribulose bisphosphate carboxylase small subunit) promoter, fused to the β-glucuronidase (GUS) reporter gene, was delivered to black spruce (Piceamariana (Mill.) BSP) tissues via microprojectile DNA bombardment, and its regulation was studied. The expression of the tobacco rbcS promoter–GUS chimeric gene was dependent on the presence of chloroplasts in black spruce tissues, as demonstrated in two ways: (i) there was no GUS activity expressed in zygotic embryos where no chloroplasts were observed, whereas it was expressed in light- and dark-grown seedlings that contained mature or immature chloroplasts; (ii) a herbicide, Norflurazon, destroyed chloroplast structure in seedlings and inhibited the expression of the tobacco rbcS promoter–GUS chimeric gene. A control chimeric gene, the cauliflower mosaic virus (CaMV) 35S promoter–GUS fusion gene was not inhibited by Norflurazon. Unlike in angiosperms, light had no effect on the expression of tobacco rbcS promoter–GUS chimeric gene. Both light- and dark-grown seedlings showed GUS activity, and expression in dark-grown seedlings was not enhanced by light. These results suggest that the tissue-specific regulation of the rbcS promoter may be conserved between angiosperms and conifers, but that the light regulation of this promoter may not be conserved.

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