Use of Anther Culture Method in Pepper Breeding: Factors Affecting Plantlets Production

Nervo, G.; Carannante, G.; Azzimonti, M. T.; Rotino, Giuseppe Leonardo

doi:10.1007/978-94-011-0307-7_20

Cited by 7 publications

(6 citation statements)

References 4 publications

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“…Beneficial effects of AgNO 3 with activated charcoal on efficiency of androgenesis in pepper anther culture have been reported (Nervo et al 1995;Comlekcioglu et al 2001;Caglar et al 2004;. Luz et al (1996Luz et al ( , 1997 observed improved embryogenic induction in medium containing only AgNO 3 , but inhibition of the processes when combined with activated charcoal.…”

Section: Plant Growth Regulatorsmentioning

confidence: 84%

Anther culture in pepper (Capsicum annuum L.) in vitro

2011

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Pepper (Capsicum annuum L.) is an important vegetable crop that can be improved using plant tissue culture and biotechnology. However, it is difficult to develop appropriate breeding material by in vitro cultivation in this species. Haploid plant production is useful in the breeding programs to facilitate recovery of recessive mutations and unique genetic recombinations. In embryogenesis, haploid formation from pollen in anther culture is a scientifically advanced, but controversial system. Various techniques for haploid plant regeneration are used to establish an efficient double haploid production method. The purpose of this article is to summarize, through comparison, results in pepper anther culture, problems associated with work in this field, and the influence of critical factors for successful embryo formation and plantlet development.

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Section: Plant Growth Regulatorsmentioning

confidence: 84%

Anther culture in pepper (Capsicum annuum L.) in vitro

2011

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“…How ever, de spite a se ries of im prove ments of the pro cess-in duc ing method de vel oped by Dumas et al (1981) con cern ing the changes in the com po si tion of cul ture me dia used at sub se quent stages of an ther cul ture (Vagera and Havranek 1985, Nervo et al 1995, Dolcet-Sanjuan et al 1997, which made it pos si ble to achieve high androgenesis ef fec tive ness of Cap si cum an num L., it is dif fi cult to trace in lit er a ture any in for ma tion on ob tain ing hap loid plants through an an ther culture of C. frutescens L. On the other hand it is rel atively easy to in duce the sporophyte de vel op ment of male gametophyte cells in a num ber of mem bers of the Solanaceae to which Capsicum ge nus belongs (Ma he shwari et al 1982). This method is frequently used in breeding of these species.…”

Section: Introductionmentioning

confidence: 99%

Induced androgenesis of Capsicum frutescens L.

2006

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De part ment of Ge net ics and Plant Breed ing, Uni ver sity of Tech nol ogy and Ag ri cul ture, Ab stractThe aim of the re search was to make a pre lim i nary de ter mi nation of the ef fec tive ness of the in duc tion of hap loids in Cap sicum frutescens L. In or der to in duce androgenesis red and yellow fruit forms of spe cies were used, each bred by the re searchers on their own. The ex per i ment was per formed in Oc to ber. An ther cul tures were con ducted ac cord ing to a mod i fied method de vel oped by Dumas et al. (1981) for C. annuum L. The an thers were laid on CP me dium con tain ing 0.01 mg·dm -3 2.4-D and 0.01 mg·dm -3 kinetin, with the ad di tion of 0.5 g·dm -3 of ac ti vated car bon and 5 mg×dm -3 of sil ver ni trate, solid i fied with 8 g·dm -3 of agar. The cul tures were in cu bated in the dark at 35 deg C for 8 days. Next they were trans ferred to 25 deg C un der a 12-hour photoperiod. Af ter 14 days of in duc tion, an thers were trans ferred to R 1 me dium sup ple mented with 0.1 mg·dm -3 kinetin. Ob tained em bryos were sub se quently transplanted onto V 3 hor mone-free me dium and well grow ing plants were planted in green houses. The ef fi ciency of androgenesis for both C. frutescens L. forms was rel a tively low and it did not ex ceed 5 %. The ploidy level of the re sult ing plants was de ter mined by flow-cytometric anal y sis. The regenerants con sisted of about equal num bers of hap loids and dip loids. Addi tion ally, among plants re gen er ated from an thers of yel low fruit forms, two mixoploids were ob served. In tro duc tionCap si cum frutescens L. plays an im por tant part among veg e ta bles, spices and med i cal plants. It is used not only in the ar eas where it orig i nated from (Carvalho et al. 2003), but also in Asia (Sreelathakumary and Rajmony 2003) and Af rica (Alegbejo and Orakwue 2002). The spe cies is a source of many in ter est ing fea tures and the fea ture-con dition ing genes. There fore it can be crossed with C. annuum L., be ing the most sig nif i cant of Cap si cum spe cies, which gives the op por tu nity of greater genetic vari a tion. The re cent re search car ried out in this re spect Olszewska 1999, Nowaczyk and Nowaczyk 2004a, b) has pointed to the prac ti ca bil ity of C. frutescens L. ge no types.One of the re cent breed ing meth ods, which have been suc cess fully used in C. annuum L. breed ing for twenty years in or der to ob tain fully homozygotic lines of dou bled hap loids (DH), is androgenesis. How ever, de spite a se ries of im prove ments of the pro cess-in duc ing method de vel oped by Dumas et al. (1981) con cern ing the changes in the com po si tion of cul ture me dia used at sub se quent stages of an ther cul ture (Vagera and Havranek 1985, Nervo et al. 1995, Dolcet-Sanjuan et al. 1997, which made it pos si ble to achieve high androgenesis ef fec tive ness of Cap si cum an num L., it is dif fi cult to trace in lit er a ture any in for ma tion on ob tain ing hap loid plants through an an ther culture of C. frutescens L. On the other hand ...

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“…Estes valores são próximos aos encontrados por Nervo et al (1994) Apesar de dois e quatro dias terem sido os melhores tempos para a indução de embrióides nas anteras e terem promovido os maiores números de embrióides, o período de oito dias foi o mais eficiente na regeneração dos embriões em plântulas (Tabela 2). Somente anteras provenientes do meio de indução C adicionado de 5 mg/L de AgNO 3 regeneraram plântulas, sendo PIX22C#31 x Linha 004 e PIX22C#21 x Ikeda, os genótipos mais responsivos (Tabela 2).…”

Section: Resultsunclassified

“…Os resultados assemelham-se aos obtidos por Nervo et al (1994), que alcançaram altas frequências de embrióides, mas baixas taxas de conversão em relação ao número de embrióides. Os autores afirmam que este é o ponto de estrangulamento da técni-ca, e sugerem maiores estudos de fatores que possam estar relacionados com tal situação.…”

Section: Resultsunclassified

“…Em pimentão, verificou-se que a presença de 5 mg/L de nitrato de prata (AgNO 3 ) no meio de indução C (Sibi et al, 1979) teve efeito favorável na androgênese, provavelmente devido ao fato do AgNO 3 bloquear a ação inibitó-ria do etileno endógeno dos embriões, permitindo assim o maior desenvolvimento dos mesmos (Nervo et al,1994). Em suspensão de células de cenoura, tanto o ácido salicílico (AS) quanto o ácido acetilsalicílico (AAS), em forma de aspirina, estimularam a embriogênese, numa faixa de concentração de 10 à 100µM, e não causaram qualquer prejuízo no crescimento e sobrevivência das células (Herman, 1991).…”

Section: Introductionunclassified

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Ação do etileno em combinação com thidiazuron, nitrato de prata e ácido acetilsalicílico na cultura de anteras de pimentão

Luz

Pinto²,

Ehlert³

et al. 1999

Hortic. Bras.

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RESUMOVerificou-se o efeito do etileno em combinação com o thidiazuron (TDZ), nitrato de prata (AgNO 3 ) e ácido acetilsalicílico (AAS), na indução e regeneração de embriões em anteras de pimentão, genótipos F 1 (PIX21C12#35 x Agronômico 08), (PIX21C04#4 x Linha 004), (PIX22C#31 x Linha 004), (PIX21C15#45 x Ikeda) e (PIX22C#21 x Ikeda). Os botões foram coletados quando sépalas e pétalas tinham tamanhos aproximadamente iguais, correspondendo ao estádio de anteras com micrósporos uninucleados. As anteras foram inoculadas em placa de Petri contendo meio de cultura C adicionado de 4,5 mM de TDZ; meio C acrescido de 0,05 mM de 2,4-D, 0,05 mM de cinetina e 88,8 mM de AAS; meio C acrescido de 0,05 mM de 2,4-D, 0,05 mM de cinetina e 5,0 mg/L de AgNO 3 . Em seguida, foram colocadas em ambiente escuro a 35 o C durante oito dias e enriquecido com ethephon, por 0; 2; 4; 6 ou 8 dias num esquema fatorial com quatro repetições, sendo cada placa considerada uma parcela contendo 20 a 24 anteras. O período de permanência de quatro dias, em ambiente enriquecido com ethephon, e os meios com TDZ e AgNO 3 , foram os mais favoráveis à indução de anteras. O TDZ também promoveu maior indução de calos embriogênicos. As maiores taxas de necrose ocorreram no meio com AAS. Só ocorreu regeneração direta em plântulas no meio C acrescido de 5 mg/L de AgNO 3 , sendo oito dias o melhor período. Os genótipos mais responsivos foram PIX22C#31 x linha 004 e PIX22C#21 x Ikeda. Observou-se a formação de plântulas haplóides e diplóides mediante análise da ponta de raíz. Considerando-se que a regeneração foi direta, sem passar por calos, supõe-se que as plântulas diplóides obtidas sejam provenientes dos micrósporos, devendo portanto ter ocorrido uma diploidização in vitro. Palavras ABSTRACTEthylene action in combination with Thidiazuron (TDZ), silver nitrate (AgNO 3 ) and acetylsalicylic acid on sweet pepper anther culture.The influence of ethylene used in combination with Thidiazuron (TDZ), silver nitrate (AgNO 3 ) and acetylsalicylic acid (ASA) was evaluated on sweet pepper androgenesis of F 1 genotypes (PIX21C12#35 x Agronômico 08, PIX21C04#4 x Line 004, PIX22C#31 x Line 004, PIX21C15#45 x Ikeda and PIX22C#21 x Ikeda). Floral buds were collected when sepals and petals were approximately of equal size, corresponding to the uninucleated stage of microspores. Anthers were placed in Petri dishes containing three mediums: C medium supplemented with TDZ (4.5mM); C medium plus 2,4-D (0,05mM), Kinetin (0.5mM) and ASA (88,8mM) or AgNO 3 (5.0 mg/L). After inoculation, the anthers were kept in the environment with ethephon for 0, 2, 4, 6 or 8 days at 35 o C for eight days in the dark. The experiment was conducted in a factorial design with four replications. Each replicate consisted of one Petri dish containing 20-24 anthers. TDZ and AgNO 3 were most effective for inducing somatic embryos after four days with Ethrel. The greatest rates of necrosis took place on the medium with AAS. Direct regeneration of plantlets occurred only on medium supplemented with 5 mg...

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Use of Anther Culture Method in Pepper Breeding: Factors Affecting Plantlets Production

Cited by 7 publications

References 4 publications

Anther culture in pepper (Capsicum annuum L.) in vitro

Anther culture in pepper (Capsicum annuum L.) in vitro

Induced androgenesis of Capsicum frutescens L.

Ação do etileno em combinação com thidiazuron, nitrato de prata e ácido acetilsalicílico na cultura de anteras de pimentão

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