scholarly journals Improved Anther Culture Media for Enhanced Callus Formation and Plant Regeneration in Rice (Oryza sativa L.)

Plants ◽  
2021 ◽  
Vol 10 (5) ◽  
pp. 839
Author(s):  
Jauhar Ali ◽  
Katrina Leslie C. Nicolas ◽  
Shahana Akther ◽  
Azerkhsh Torabi ◽  
Ali Akbar Ebadi ◽  
...  

Anther culture technique is the most viable and efficient method of producing homozygous doubled haploid plants within a short period. However, the practical application of this technology in rice improvement is still limited by various factors that influence culture efficiency. The present study was conducted to determine the effects of two improved anther culture media, Ali-1 (A1) and Ali-2 (A2), a modified N6 medium, to enhance the callus formation and plant regeneration of japonica, indica, and hybrids of indica and japonica cross. The current study demonstrated that genotype and media had a significant impact (p < 0.001) on both callus induction frequency and green plantlet regeneration efficiency. The use of the A1 and A2 medium significantly enhanced callus induction frequency of japonica rice type, Nipponbare, and the hybrids of indica × japonica cross (CXY6, CXY24, and Y2) but not the indica rice type, NSIC Rc480. However, the A1 medium is found superior to the N6 medium as it significantly improved the green plantlet regeneration efficiency of CXY6, CXY24, and Y2 by almost 36%, 118%, and 277%, respectively. Furthermore, it substantially reduced the albino plantlet regeneration of the induced callus in two hybrids (CXY6 and Y2). Therefore, the improved anther culture medium A1 can produce doubled haploid rice plants for indica × japonica, which can be useful in different breeding programs that will enable the speedy development of rice varieties for resource-poor farmers.

2016 ◽  
Vol 44 (2) ◽  
pp. 133
Author(s):  
Cucu Gunarsih ◽  
Bambang Sapta Purwoko ◽  
Iswari Saraswati Dewi ◽  
Dan Muhamad Syukur

ABSTRACT<br /><br />The breeding of rainfed rice tolerant to drought can be accomplished using anther culture. The objectives of this research were to determine regeneration abilities of six F1 anther culture and its acclimatization ability. The experiment was arranged in completely randomized design with 14 replications. The treatments consisted of six F1 derived from crossing:  INPARI 18 x IR83140-B-11-B (G1), INPARI 18 x B12825E-TB-1-25 (G2), INPARI 18 x IR87705-14-11-B-SKI-12 (G3), INPARI 22 x IR83140-B-11-B (G4), Bio-R81 x O18b-1 (G5), Bio-R82-2 x O18b-1 (G6). Media for callus induction was based on N6 medium + 2.0 mg L-1 NAA + 0.5 mg L-1 kinetin + 1.0 mM putresin + 60 g L-1 sucrosa, media for regeneration was based on MS + 0.5 mg L-1 NAA + 2.0 mg L-1 kinetin + 1.0 mM  putresin, and media for rooting was based on  MS + 0.5 mg L-1 IBA + 30 g L-1 sucrosa. The result indicated that all six F1 had different ability in anther culture. Bio-R82-2 x O18-b1 (G6) and  Bio-R81 x O18-b1 (G5) F1 genotype had good response both of callus induction and plant regeneration. These two F1 genotypes also gave the highest ratio of green planlet production to number of anther inoculated (GP:AI) were 5.50% and 4.65%,  respectively. In this research, there were identified doubled haploid plants were developed from 4 F1 derived cross namely G2 (2 plants), G3 (4 plants),  G5 (21 plants), and G6 (26 plants).<br /><br />Keywords: Callus induction, doubled haploid, rice<br /><br />


2012 ◽  
Vol 12 (2) ◽  
pp. 93 ◽  
Author(s):  
Budi Winarto ◽  
Nurhayati Ansori Mattjik ◽  
Agus Purwito ◽  
Budi Marwoto

Improvement of selected induction culture media on callus induction in anther culture of anthurium and a histologicalstudy on its callus formation were studied at the tissue culture laboratory of the Indonesian Ornamental CropsResearch Institute from February to October 2008. The objectives of the study were to optimize selected media forcallus formation, reveal cell origin of callus derived from anther culture and shoot formation process. Selectedmedia improved in the study were 1) MMS-TBN containing 0,5 mg/l TDZ, 1,0 mg/l BAP and 0,01 mg/l NAA (Winartomedium, WM) and 2) MMS III supplemented with 1,5 mg/l TDZ, 0,75 mg/l BAP and 0,02 mg/l NAA (Winarto andRachmawati medium, WRM). Improvement treatments were carried out by omission and application of 2,4-D in 0.5mg/l and reduction of medium strength of full, half, quarter, one eighth, one sixteenth, and zero strength. Afactorial experiment was arranged using a randomized complete block design with four replications. Results ofthis study indicated that the highest callus induction was clearly established in WRM. The medium stimulatedpotential growth of anther (PGA) up to 81% with 49% of percentage of anther regeneration (PAR) and 2.7 number ofcallus formed per replication (NCF). Significant improvement in callus formation was also recorded by reduction ofmedium strength of WRM to one eighth compared to others. The reduction induced PGA up to 58% with 29% of PARand 1.8 NCF. From histological studies it was well recognized that regenerated callus on half anthers cultured wasoriginated from middle layer cells of anther wall. The morphogenic response of anther wall cells caused primarilyon no androgenesis effect in microspore cells.


1970 ◽  
Vol 34 (4) ◽  
pp. 693-703 ◽  
Author(s):  
MA Alam ◽  
MA Haque ◽  
MR Hossain ◽  
SC Sarker ◽  
R Afroz

Anther of five varieties of Brassica species, namely BARI Shariaha-7, Tori-7, Agrani, Daulat and Safal were cultured in vitro to observe their regeneration potentiality. Different concentrations and combinations of growth regulators were supplemented in MS medium. The range of callus induction was 12.50-87.50 %. Maximum callus induction (75.00%) was observed on MS +4 mg/L 2, 4-D + 1.0 mg/L BAP. Among the genotypes, BARI Sharisha-7 showed the highest percentage of callus induction (60.42%). Among the treatments, highest percentage of shoot regeneration (75.00%) was observed on MS + 4 mg/L BAP + 1.0 mg/L NAA. BARI Sharisha-7 also showed the highest rate of plant regeneration (66.67%). Root induction was highest (75%) on half strength MS medium supplemented with 1.0 mg/L IBA and 0.5 mg/L NAA. The plantlets with sufficient roots thus obtained were transferred successfully to plastic pots and subsequently to the field. BARI Sharisha-7 and Tori-7 survived easily in the pots as well as in the field but Safal was very poor in survivability both in the pots and in the field. Key Words: Brassica; haploid; anther culture; in vitro regeneration.DOI: 10.3329/bjar.v34i4.5844Bangladesh J. Agril. Res. 34(4) : 693-703, December 2009 


2016 ◽  
Vol 9 (2) ◽  
pp. 60 ◽  
Author(s):  
Iswari S. Dewi ◽  
Bambang S. Purwoko

The polyamines such as putrescine, spermidine, and spermine were reported to increase green plant regeneration in rice anther culture. Low response of anther culture of rice sub-species indica may be improved with the addition of putrescine in the culture media. Four experiments were conducted to study the role of polyamines in inhibition of ethylene biosynthesis and their effects on rice anther culture development. Anthers of two subspecies of rice, indica (IR64, Krowal, Jatiluhur) and japonica (Taipei 309) were cultured onto media supplemented with putrescine (N6P) and without putrescine (N6). Young<br />panicles containing the anthers at mid-to-late nucleate microspores were cold pretreated at 5 + 2°C and incubated in the dark for 8 days before the anthers were cultured. Results<br />showed that medium without putrescine produced an earlier senescence of indica rice anther than that of japonica. The addition of 10-3 M putrescine into the culture media inhibited ethylene biosynthesis as anther senescence delayed, increased the three polyamines contents, and decreased the ACC content as well as ACC oxydase activity in anther-derived calli. In the anther and anther-derived calli of subspecies indica, the total<br />polyamines content was lower (10.14 nM g-1 anther and 8.48 nM g-1 calli) than that of subspecies japonica (12.61 nM g-1 anther and 10.16 nM g-1 calli), whereas the ethylene production was higher (32.31 nM g-1 anther and 2.48 nM g-1 calli) than the japonica (31.68 nM g-1 anther and 1.76 nM g-1 calli). This study suggests that application of 10-3 M putrescine in anther culture of rice subspecies indica improves androgenesis by inhibiting<br />early senescence of cultured anthers and enhancing embryo or callus formation from microspores.


2013 ◽  
Vol 18 (2) ◽  
pp. 1-9
Author(s):  
MAZ Al Munsur ◽  
MS Haque ◽  
KM Nasiruddin ◽  
MJ Hasan

Experiments were carried out to study the callus induction and subsequent plantlet regeneration ability of leaf segments and root tips of bitter gourd. A combination of 1.0 mg L-1 2,4-D and 1.0 mg L-1 BAP produced the highest percentage of callus in leaf segments (78.75%) and 1.0 mg L-1 2,4-D combined with 0.5 mg L-1 BAP produced the highest callus frequency in root tips (72.5%). Using the combination of BAP at 2.0 mg L-1 and NAA at 0.3 mg L-1 improved the callus induction frequency (80.00%) in root tips. Root tips showed better response than leaf segments in callus induction. Leaf segments and root tips had equal percentage (65.0) of shoot regeneration upon culture of the calli on medium with 2.0 or 2.5 mg L-1 BAP with 0.2 mg L-1 IAA. The highest length (3.95 cm) of shoot from leaf segments was recorded with 2.5 mg L-1 BAP, 0.5 mg L-1 IBA and 0.2 mg L-1 GA3. The highest number of roots (6.75) and root length (2.45 cm) were found in 3.0 mg L-1 BAP with 0.1 mg L-1 NAA in root tips. The callus formation and regeneration ability of the explants have the potential for use in future biochemical studies and genetic improvement of bitter gourd.DOI: http://dx.doi.org/10.3329/pa.v18i2.17457 Progress. Agric. 18(2): 1 - 9, 2007


Author(s):  
Mridhu Sharma ◽  
A. K. Singh ◽  
R. K. Salgotra ◽  
Manmohan Sharma ◽  
Bupesh Ku. Sharma ◽  
...  

With different culture conditions and concentrations of growth regulators and supplements, Anther culture technique can be easily employed for the production of haploids under in vitro conditions. Aims: The present study was undertaken with the objective to optimize the development of doubled haploids using anthers for in vitro induction of callus on N6 medium. Place and Duration of Study: The samples (BC2F1 seeds) were raised previously in Skuast-J. From total degree program of 3 years, this work related to tissue culture technique was done in one year from January 2018 to January 2019. Methodology: The effect of levels of 2, 4-dichlorophenoxy acetic acid (2, 4-D) i.e. 0 to 3 mg/L in basal N6 media was observed on callus induction frequency (CIF). The effect of duration of cold pre-treatment was observed on callus induction frequency at 2.5 mg/L of 2, 4-D by giving the cold pre-treatment at 4ºC from 8 to 12 days. Also the effect of different amino acids was checked on callus induction frequency. Results: Highest callus induction frequency of 9.39 per cent was observed in N6 medium fortified with 2.5 mg/L 2, 4-D and lowest callus induction frequency of 2.52 per cent at the concentration of 1.0 mg/L. The cold pre-treatment for 10 days gave highest callus induction frequency of 1.44 per cent and lowest callus induction frequency of 0.44 per cent was obtained at cold pre treatment for 8 days. The highest callus induction frequency of 12.55 per cent was observed in case of media supplemented with 25 mg/L tryptophan and 40 mg/L cysteine and lowest callus induction frequency of 7.18 per cent was observed when media was supplemented with 560 mg/L proline. Conclusion: The cold pre-treatment of 10 days at 4ºC on media supplemented with 2.5 mg/L of 2, 4-D and combination of 25 mg/L tryptophan and 40 mg/L cysteine proves to provide best androgenesis conditions for anthers from BC2F1 population.


Author(s):  
RK Niroula ◽  
BP Sah ◽  
HP Bimb ◽  
S Nayak

This study was under taken to elucidate the effect of genotypes and media compositions on callus induction from mature rice seeds. Three different callus induction media, designated as A (N6 mineral salts + N6 vitamins, 2 mg/l each + myoinositol, 100 mg/l + 2,4-D, 2.5 mg/l + kinetin, 0.5mg/l + AgNO3, 10 mg/l + maltose, 50 gm/l); B (MS organic salts + N6 mineral salts + NAA, 4 mg/l + kinetin, 2 mg/l + AgNO3, 5mg/l and sucrose, 60 gm/l); and C (B media without AgNO3), and six rice genotypes viz. Jumlimarshi, Tilki, Jethobudo, Manshara, Masuli and Pahenle were evaluated. The modified N6 medium supplemented with 2, 4-D, 2.5 mg/l and AgNO3, 10 mg/l exhibited better performance in callus induction. Among genotypes, callus induction frequency was higher (100%) in Masuli, Tilki and Jumlimarshi regardless of media tested. The positive effect of AgNO3 was only observed in medium A for quality callus induction and subsequent plant regeneration. The genotype Tilki performed better regarding plant regeneration (27.77%). Therefore, it is suggested that application of medium A is advantageous to accomplish overall efficiency of callus induction and plant regeneration from seeds of various rice genotypes. Key words: 2, 4-D; AgNO3; dehulled rice; in vitro; regeneration J. Inst. Agric. Anim. Sci. 26:21-26 (2005)


2016 ◽  
Vol 9 (2) ◽  
pp. 60 ◽  
Author(s):  
Iswari S. Dewi ◽  
Bambang S. Purwoko

The polyamines such as putrescine, spermidine, and spermine were reported to increase green plant regeneration in rice anther culture. Low response of anther culture of rice sub-species indica may be improved with the addition of putrescine in the culture media. Four experiments were conducted to study the role of polyamines in inhibition of ethylene biosynthesis and their effects on rice anther culture development. Anthers of two subspecies of rice, indica (IR64, Krowal, Jatiluhur) and japonica (Taipei 309) were cultured onto media supplemented with putrescine (N6P) and without putrescine (N6). Young<br />panicles containing the anthers at mid-to-late nucleate microspores were cold pretreated at 5 + 2°C and incubated in the dark for 8 days before the anthers were cultured. Results<br />showed that medium without putrescine produced an earlier senescence of indica rice anther than that of japonica. The addition of 10-3 M putrescine into the culture media inhibited ethylene biosynthesis as anther senescence delayed, increased the three polyamines contents, and decreased the ACC content as well as ACC oxydase activity in anther-derived calli. In the anther and anther-derived calli of subspecies indica, the total<br />polyamines content was lower (10.14 nM g-1 anther and 8.48 nM g-1 calli) than that of subspecies japonica (12.61 nM g-1 anther and 10.16 nM g-1 calli), whereas the ethylene production was higher (32.31 nM g-1 anther and 2.48 nM g-1 calli) than the japonica (31.68 nM g-1 anther and 1.76 nM g-1 calli). This study suggests that application of 10-3 M putrescine in anther culture of rice subspecies indica improves androgenesis by inhibiting<br />early senescence of cultured anthers and enhancing embryo or callus formation from microspores.


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


2004 ◽  
Vol 7 (3) ◽  
pp. 331-334 ◽  
Author(s):  
Md. Monirul Islam ◽  
Sanjay Kumar Adhikar . ◽  
Purnendu Gain . ◽  
Md. Mizanur Rahman . ◽  
Noor-e-Alam Siddique .

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