scholarly journals Plant regeneration from in vitro leaves of four commercial Pyrus species

2008 ◽  
Vol 54 (No. 4) ◽  
pp. 140-148 ◽  
Author(s):  
H. Tang ◽  
Y. Luo ◽  
C. Liu

An efficient shoot regeneration from in vitro leaf sections of <I>Pyrus communis</I> Bartlett, <I>P. pyrifolia</I> Shenbuzhi, <I>P. bretschneideri</I> Zaosu and <I>P. ussuriensis</I> Manyuanxiang was successfully developed for use in future transgenic studies. On the basis of regeneration frequency and average shoot numbers, optimal shoot regeneration was obtained on leaf sections of <I>P. communis</I> Bartlett when cultured on Murashige and Skoog complete medium containing 6.0 mg/l BA (6-benzyladenine) and 0.1 mg/l NAA (&alpha;-naphthaleneacetic acid), while Quoirin and Lepoivre complete medium supplemented with 1.0 mg/l TDZ [thidiazuron (N-phenyl-N<sup>1</sup>-1,2,3-thiadiazol-5-ylurea)] and 0.1 mg/l NAA was found best for <I>P. pyrifolia</I> Shenbuzhi, and Nitsch and Nitsch complete medium containing 3.0 mg/l TDZ and 0.1 mg/l NAA or 0.2 mg/l IAA was suitable for<I>P. bretschneideri</I> Zaosu or <I>P. ussuriensis</I> Manyuanxiang, respectively. After cutting the leaves into three sections perpendicular to the midrib and culturing under the equivalent conditions, regeneration occurred more frequently on basal sections than middle sections, and no shoots formed on apical sections. A ratio of NH<sup>+</sup><sub>4</sub>-N/NO<sup>-</sup><sub>3</sub>- N of 1:2~1:7 was found beneficial for shoot regeneration. 75.0–87.5% of proliferating shoots formed roots after 4 weeks of transfer to 1/4 strength of Murashige and Skoog complete medium supplemented with 2.5 mg/l IBA (indole-3-butryric acid) and 30.0 g/l sucrose. Regenerated plants were successfully established under greenhouse conditions.

2020 ◽  
Vol 10 (15) ◽  
pp. 5370 ◽  
Author(s):  
Mohsen Hesami ◽  
Jorge A. Condori-Apfata ◽  
Maria Valderrama Valencia ◽  
Mohsen Mohammadi

Optimizing in vitro shoot regeneration conditions in wheat is one of the important steps in successful micropropagation and gene transformation. Various factors such as genotypes, explants, and phytohormones affect in vitro regeneration of wheat, hindering the ability to tailor genotype-independent protocols. Novel computational approaches such as artificial neural networks (ANNs) can facilitate modeling and predicting outcomes of tissue culture experiments and thereby reduce large experimental treatments and combinations. In this study, generalized regression neural network (GRNN) were used to model and forecast in vitro shoot regeneration outcomes of wheat on the basis of 10 factors including genotypes, explants, and different concentrations of 6-benzylaminopurine (BAP), kinetin (Kin), 2,4-dichlorophenoxyacetic acid (2,4-D), indole-3-acetic acid (IAA), indole-3-butyric acid (IBA), 1-naphthaleneacetic acid (NAA), zeatin, and CuSO4. In addition, GRNN was linked to a genetic algorithm (GA) to identify an optimized solution for maximum shoot regeneration. Results indicated that GRNN could accurately predict the shoot regeneration frequency in the validation set with a coefficient determination of 0.78. Sensitivity analysis demonstrated that shoot regeneration frequency was more sensitive to variables in the order of 2,4-D > explant > genotype < zeatin < NAA. Results of this study suggest that GRNN-GA can be used as a tool, besides experimental approaches, to develop and optimize in vitro genotype-independent regeneration protocols.


2010 ◽  
Vol 40 (10) ◽  
pp. 2210-2213
Author(s):  
Monalize Salete Mota ◽  
Juliana de Magalhães Bandeira ◽  
Eugenia Jacira Bolacel Braga ◽  
Valmor João Bianchi ◽  
José Antonio Peters

A shoot regeneration system for Plectranthus neochilus was studied from leaf explants. Leaves developed under in vitro conditions were cultured on Wood Plant Medium supplemented with 0.2mg dm-3 α-naphthaleneacetic acid (NAA) and different 6-benzilaminopurine (BAP) or thidiazuron (TDZ) concentrations (0, 1.5, 3.0, 4.5 and 6.0mg dm-3). An increase in percentage of responsive explants (85.3%) and in the number of shoots developed per explant (3.2) was observed when the explants were treated with 5.3 and 4.7mg dm-3 BAP, respectively. The leaf explants cultured on media supplemented with TDZ became vitreous and did not form buds. The regeneration system used is efficient for boldo bud induction and shoot development, showing high potential for advanced cellular and molecular studies.


1970 ◽  
Vol 35 (2) ◽  
pp. 331-341 ◽  
Author(s):  
MA Sayem ◽  
M Maniruzzaman ◽  
SS Siddique ◽  
M Al-Amin

The experiment was conducted to investigate the performance of three different genotypes (BARI Sarisha-6, BARI Sarisha-8, and BARI Sarisha-11) in two different media viz., MS and B5 with different concentrations of phytohormone (2, 4-D) for callus induction from uninucleate stage anthers of Brassica and subsequent plant regeneration in MS media with different concentrations of phytohormone (BAP and NAA). Among the genotypes, BARI Sarisha-8 showed the best performance for all the parameters of callus induction. The performance of BARI Sarisha-6 was poor compared to others. Maximum rate of callus induction (%) was observed in MS + 0.5 mg/L 2, 4-D followed by B5 + 0.5 mg/L 2,4-D. The media combination MS + 1.0 mg/L BAP 0.3 mg/L 2,4-D showed the best performance for maintenance of calli. Significant variations were observed among the genotypes and media composition for shoot regeneration. Among the genotypes, BARI Sarisha-8 showed the best performance for shoot regeneration followed by BARJ Sarisha-l1. The genotype BARI Sarisha-8 produced higher percent of shoots/calli and required minimum days for shoot initiation. Higher percent calli without shoot were produced by the genotype BARI Sarisha-6. The media combination MS + 2.0 mg/L BAP + 0.5 mg/L NAA showed the best performance for shoot regeneration and required maximum days for shoot initiation. Keywords: Regeneration; BARI Sarisha-6; BARI Sarisha-8; BARI Sarisha-11; anther culture; phytohormone  DOI: 10.3329/bjar.v35i2.5896Bangladesh J. Agril. Res. 35(2) : 331-341, June 2010


2008 ◽  
Vol 43 (10) ◽  
pp. 1325-1330 ◽  
Author(s):  
Lucymeire Souza Morais-Lino ◽  
Janay Almeida dos Santos-Serejo ◽  
Sebastião de Oliveira e Silva ◽  
José Raniere Ferreira de Santana ◽  
Adilson Kenji Kobayashi

The objective of this study was to establish cell suspension culture and plant regeneration via somatic embryogenesis of a Brazilian plantain, cultivar Terra Maranhão, AAB. Immature male flowers were used as explant source for generating highly embryogenic cultures 45 days after inoculation, which were used for establishment of cell suspension culture and multiplication of secondary somatic embryos. Five semisolid culture media were tested for differentiation, maturation, somatic embryos germination and for plant regeneration. An average of 558 plants per one milliliter of 5% SCV (settled cell volume) were regenerated in the MS medium, with 11.4 µM indolacetic acid and 2.2 µM 6-benzylaminopurine. Regenerated plants showed a normal development, and no visible somaclonal variation was observed in vitro. It is possible to regenerate plants from cell suspensions of plantain banana cultivar Terra using MS medium supplemented with 11.4 µM of IAA and 2.2 µM of BAP.


2021 ◽  
Author(s):  
Yuan-yuan Meng ◽  
Shi-jie Song ◽  
Sven Landrein

Abstract Passiflora xishuangbannaensis (Passifloraceae) is endemic to a few sites of Mengyang nature reserve in Yunnan, Xishuangbanna and less than 40 individuals have been recorded. Nine Passiflora species are endemic to Yunnan with most species occurring in South America, making P. xishuangbannaensis highly significant and emblematic to the conservation work in the region. This study is designed to provide the first protocol for in vitro organogenesis and plant regeneration for ex situ conservation and reintroduction for an Asian Passiflora species. Using internodes, petioles and tendrils we optimize calli formation and root elongation using several plant growth regulators, individually or in combination. We also assess the genetic stability of regenerated cells. The maximum callus induction and shoot bud differentiation were both achieved on half Murashige and Skoog basal medium supplemented with 4.44 µM 6-Benzylaminopurine and 1.08 µM 1-Naphthaleneacetic acid. The best rooting was achieved from 30 days old, regenerated shoots on half Murashige and Skoog basal medium supplemented with 1.08 µM 1-Naphthaleneacetic acid. Micropropagated plants were subjected to inter simple sequence repeat markers analyses. Collectively, 86 bands were generated from 6 primers of which 12 bands were polymorphic, showing genetic variation between the regenerated plantlets and the original plant. Response to plant growth regulators was more specific than most other studies using South American species, which could be explained by the morphological and physiological differences between South American and Asian Passiflora species


2018 ◽  
Vol 46 (1) ◽  
pp. 52-57 ◽  
Author(s):  
Alina TREJGELL ◽  
Monika KAMIŃSKA ◽  
Karolina LISOWSKA ◽  
Andrzej TRETYN

This is the first communication of micropropagation system for Inula germanica using seedling explants germinated in vitro. The development of this system gives the possibility of future reintroduction of I. germanica providing a way to stabilize or re-establish its population. Shoot tips and fragments of cotyledons, hypocotyls and roots were isolated from ten-day-old seedlings. Explants were put on MS medium containing 1.0 mg l-1 benzylaminopurine and 0.1 mg l-1 naphthaleneacetic acid and cultured under continuous white fluorescent light (45 μmol.m-2.s-1) at 26 ± 1 °C. The highest percentage of shoot organogenesis (83.3%) was recorded for hypocotyl, while the highest average number of shoots per explant (12.0) was recorded for shoot tips. In subsequent subcultures, multiplication rate decreased to 3.0-4.9 shoots per explant. Less than 19% shoots were able to root on the solid medium without auxins. The highest rooting efficiency (69.3%) was recorded for solid medium supplemented with indolebutyric acid, but growth of roots was inhibited. The percentage of rooted shoots (62.2%) and number of roots per shoot (2.4 per shoot) into the liquid medium were comparable to medium with 0.1 mg·l-1 indolebutyric acid. showing a positive impact on the process of acclimatization. The regenerated plants were able to flowering in the first year after acclimatization. Developed micropropagation system for I. germanica is efficient and can be a useful tool for the active protection of this species.


Plants ◽  
2020 ◽  
Vol 9 (3) ◽  
pp. 318
Author(s):  
Mehtab Muhammad Aslam ◽  
Joseph K. Karanja ◽  
Qian Zhang ◽  
Huifeng Lin ◽  
Tianyu Xia ◽  
...  

The tissue culture regeneration system of Lupinus albus has always been considered as recalcitrant material due to its genotype-dependent response and low regeneration efficiency that hamper the use of genetic engineering. Establishment of repeatable plant regeneration protocol is a prerequisite tool for successful application of genetic engineering. This aim of this study was to develop standardized, efficient protocol for successful shoot induction from cotyledonary node of white lupin. In this study, 5 day old aseptically cultured seedlings were used to prepare three explants (half cotyledonary node, HCN; whole cotyledonary node, WCN; and traditional cotyledonary node, TCN), cultured on four concentrations of M519 medium (M519, ½ M519, 1/3 M519, and ¼ M519), containing four carbohydrate sources (sucrose, fructose, maltose, and glucose), and stimulated with various combinations of KT (kinetin), and NAA (naphthalene acetic acid) for direct shoot regeneration. High frequency of 80% shoot regeneration was obtained on ½ M519 medium (KT 4.0 mg L−1 + NAA 0.1 mg L−1) by using HCN as an explant. Interestingly, combinations of (KT 4.0 mg L−1 + NAA 0.1 mg L−1 + BAP 1.67 mg L−1), and (KT 2.0 mg L−1 + NAA 0.1 mg L−1) showed similar shoot regeneration frequency of 60%. Augmentation of 0.25 g L−1 activated charcoal (AC) not only reduced browning effect but also improved shoot elongation. Among the all carbohydrate sources, sucrose showed the highest regeneration frequency with HCN. Additionally, 80% rooting frequency was recorded on ½ M519 containing IAA 1.0 mg L−1 + KT 0.1 mg L−1 (indole acetic acid) after 28 days of culturing. The present study describes establishment of an efficient and successful protocol for direct plant regeneration of white lupin from different cotyledonary nodes.


Plants ◽  
2019 ◽  
Vol 8 (6) ◽  
pp. 178 ◽  
Author(s):  
Sabbadini ◽  
Ricci ◽  
Limera ◽  
Baldoni ◽  
Capriotti ◽  
...  

Prunus spp. is one of the most recalcitrant fruit tree species in terms of in vitro regeneration and transformation, mostly when mature tissues are used as explants. The present study describes the in vitro regeneration via indirect organogenesis, and Agrobacterium tumefaciens-mediated transformation of the peach rootstock Hansen 536 (Prunus persica × Prunus amygdalus) through the use of meristematic bulks (MBs) as starting explants. Efficient adventitious shoot regeneration was obtained when Hansen 536 MBs were cultured on an optimized medium consisting of modified McCown Woody Plant medium (WPM) enriched with 4.4 M 6-Benzyladenine (BA), 0.1 M 1-Naphthaleneacetic acid (NAA) and 6.0 g L−1 plant agar S1000 (B&V). MB slices were used later as starting explants for Agrobacterium-mediated transformation to introduce an RNAi construct “ihp35S-PPV194” against PPV virus. Transgenic events were identified by both green fluorescent protein (GFP) screening and kanamycin selection at different concentrations (0, 17 or 42 M). GFP-fluorescent proliferating callus lines were selected and confirmed to stably express the ihp35S-PPV194::eGFP gene construct by molecular analysis. Although shoot regeneration from these transgenic calli has not been obtained yet, this represents one of the few examples of successful attempts in peach genetic transformation from somatic tissues, and also serves as a useful in vitro system for future gene functional analysis in peach.


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