Embryonic axes excised from seeds ofGenipa americanaL. desiccated to different water contents were successfully cryopreserved by rapidly plunging seed samples directly into liquid nitrogen. Control and cryopreserved embryonic axes were excised and grown in WPM culture medium for viability assessment. All control embryonic axes (−LN2) excised from fully hydrated seeds (43.89% moisture content) germinated after 21 days of culturein vitro. These high germination percentages persisted even after the water content of the seeds was as low as 6.79%. After freezing in liquid nitrogen high germination percentages, 93%, 96%, and 93%, were observed for embryonic axes excised from seeds dehydrated to 13.26%, 9.57%, and 6.79 moisture content, respectively. The cryopreservation technique described here is recommended for long term conservation ofG. americanagermplasm.
Jatropha curcas L. is a plant species with many potential applications, especially medicinal uses (hypoglycemic, anti-inflammatory, haemostatic, healing, anti-tumor). The objective of this study was to test germination in moist paper rolls for whole seeds and in vitro for excised embryonic axes, in an attempt to identify the best method to assess the quality of J. curcas seed germplasm, cryopreserved with different water contents. The experimental sample with a 6.2% moisture content (MC) was divided in subsamples which were hydrated and dehydrated for 0 (control), 4, 8, 11 and 24h. The initial germination percentages were 63% for whole seeds and 81% for excised embryonic axes. After exposure to liquid nitrogen (LN), germination percentages were 48% (whole seeds) and 57% (excised embryonic axes). There was no significant difference between germination percentages in embryonic excised from seeds subjected or not subjected to freezing, with different MC. In contrast, there was a reduction of the whole seed germination percentage when exposed to LN (contrast = 0.17, standard error = 0.04, t = 4.09, p = 0.001) and not for the hydration and dehydration treatments. The methodology based on in vitro cultures of the embryonic axis isolated from seeds stored in LN with distinct MC values was more efficient than the standard germination test to evaluate the viability of J. curcas seeds before and after LN storage.Keywords: seed, embryonic axis, cryopreservation, Jatropha curcas RESUMO: Métodos para avaliar a viabilidade de germoplasma semente criopreservado de Jatropha curcas L. Jatropha curcas L., é uma espécie com várias aplicações potenciais, principalmente para usos medicinais (hipoglicemina, anti-inflamatório, homeostático, cicatrizante, antitumoral). O objetivo deste estudo foi testar a germinação em rolo de papel para sementes inteiras e in vitro para eixos embrionários excisados visando identificar o melhor método para avaliar a qualidade de germoplasma semente de J. curcas, criopreservado com diferentes teores de água. A amostra experimental com 6,2% de teor de água foi dividida em subamostras que foram hidratadas e desidratadas por 0 (controle), 4, 8, 11 e 24 h. Os percentuais de germinação inicial foram de 63% para sementes inteiras e de 81% para eixos embrionários excisados. Após exposição ao nitrogênio líquido (NL) os percentuais de germinação foram de 48% (sementes inteiras) e 57% (eixos embrionários). Não houve diferença significativa entre os percentuais de germinação de eixos embrionários excisados de sementes com diferentes umidades e submetidas ou não ao congelamento. Em contraste, houve redução de percentuais de germinação das sementes inteiras expostas ao NL (contraste = 0.17, erro padrão = 0.04, t = 4.09, p = 0.001), mas não aos tratamentos de hidratação e desidratação. A metodologia baseada em cultura de eixos embrionários in vitro isolados de sementes armazenadas em NL, com distintos teores de água foi mais eficiente que a germinação padrão para avaliar a viabilidade de sementes J. curcas ...
Queen palm (Syagrus romanzoffiana [Cham.] Glassman) is a palm species best known as an ornamental tree for urban landscaping, but recently, it has been evaluated as a potential crop for biofuel production. The objective of the present work was to establish a cryopreservation technique for queen palm to ensure long-term conservation of this species. The cryopreservation protocol consisted of direct immersion in liquid nitrogen (LN) of whole endocarps with water contents ranging from 5.5 to 10.9%, followed by slow thawing at room temperature (25 ± 2°C) excision and in vitro culture of zygotic embryos. Viability of zygotic embryos isolated from endocarps with different water contents was evaluated before (control) and after freezing in LN using in vitro culture on Woody Plant Medium (WPM) medium. Germination percentages of zygotic embryos isolated from endocarps stored in LN varied from 84 to 93%, whereas those isolated from controls ranged from 55 to 71%. Germination rates were significantly higher for zygotic embryos excised from cryopreserved endocarps. The water content of control or frozen endocarps did not have a significant effect on germination percentages of zygotic embryos. Zygotic embryos excised from endocarps following cryopreservation in liquid nitrogen developed into normal plantlets after in vitro culture. The technique tested is simple, efficient, and can be used in plant gene banks as a routine approach for long-term conservation of queen palm germplasm.
Jatropha seeds are classified as orthodox. However, since it is an oil seed species, adequate storage conditions are required to ensure their longevity. The objective of this work was to evaluate the physiological quality of jatropha seeds stored in different environments and packaging, for periods of 3, 9 and 15 months. Three types of seed packaging bags (high density plastic bag, aluminized envelope and multiwall paper bag) were used, and the storage environments were cold and dry chamber (20 °C and 15% RH, constant), refrigerator (7 ± 3 °C, 48 ± 8% RH) and laboratory conditions (25 ± 3 °C, 51 ± 7% RH). The initial moisture content and seed germination were 7.1% and 89%, respectively. During storage, the physiological quality (germination and vigor) and moisture content of the seeds were evaluated. Seed water content ranged from 3.3 to 7.7%, depending on the permeability of the packaging and the storage environment. The highest longevity (15 months) without loss of viability was observed for jatropha seeds with initial moisture of 7.1%, packed in semipermeable plastic. Seed vigor was maintained, regardless of the environment and the type of packaging used, for up to nine months of storage.
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