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Reprogramming of cassava (Manihot esculenta) microspores towards sporophytic development.

Perera PI, Ordoñez CA, Dedicova B, Ortega PE - AoB Plants (2014)

Bottom Line: In two cassava genotypes, different developmental stages of microspores were found to initiate sporophytic cell divisions, that is, with tetrads of TMS 60444 and with mid or late uni-nucleate microspores of SM 1219-9.A low frequency of the reprogramming and the presence of non-responsive microspores among the responsive ones in tetrads were found to be related to the viability and exine formation of the microspores.This paves the way for the development of an efficient technique for the production of homozygous lines in cassava.

View Article: PubMed Central - PubMed

Affiliation: Agrobiodiversity Research Area, International Center for Tropical Agriculture, A.A. 6713, Cali, Colombia Present address: Department of Horticulture and Landscape Gardening, Faculty of Agriculture and Plantation Management, Wayamba University, Gonawila, Makandura, Sri Lanka prasanthi1970@yahoo.com.

No MeSH data available.


Related in: MedlinePlus

Quantitative and qualitative demonstration of the microspore viability of tetrads of M. esculenta genotype TMS 60444. Fluorescein-diacetate-stained microspores of tetrads under (A) a light microscope and (B) a fluorescent microscope. (C) Microspores of tetrads stained with Alexander stain. Lane 1: tetrads with four viable microspores; Lane 2: with three viable microspores; Lane 3: with two viable microspores; Lane 4: with one viable microspore; Lane 5: without any viable microspores in the tetrads (scale bars = 20 µm). (D and E) Quantitative data of microspore viability in tetrads after sterilization (AS), 3 days after cold pretreatment (ACP), 3 days after culture (3DC) and 7 days after culture (7DC) as compared with fresh (F) tetrads with FDA (D) and Alexander staining (E).
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PLU022F5: Quantitative and qualitative demonstration of the microspore viability of tetrads of M. esculenta genotype TMS 60444. Fluorescein-diacetate-stained microspores of tetrads under (A) a light microscope and (B) a fluorescent microscope. (C) Microspores of tetrads stained with Alexander stain. Lane 1: tetrads with four viable microspores; Lane 2: with three viable microspores; Lane 3: with two viable microspores; Lane 4: with one viable microspore; Lane 5: without any viable microspores in the tetrads (scale bars = 20 µm). (D and E) Quantitative data of microspore viability in tetrads after sterilization (AS), 3 days after cold pretreatment (ACP), 3 days after culture (3DC) and 7 days after culture (7DC) as compared with fresh (F) tetrads with FDA (D) and Alexander staining (E).

Mentions: In order to find the reason for the low response in TMS, first the viability of the microspores in tetrads was evaluated by FDA and Alexander staining. The viability of the microspores in tetrads at the time of bud collection was compared with the viability at the different steps of culture protocol. Either all four microspores (Fig. 5, Lane 1) or three (Fig. 5, Lane 2), two (Fig. 5, Lane 3) or one microspore (Fig. 5, Lane 4) were viable in the tetrads when tested with FDA (Fig. 5A–C) in frequencies of 51.3, 25.2, 13.1 and 6 % in the freshly collected buds, whereas the frequency of tetrads with all four non-viable microspores (Fig. 5, Lane 5) was 4.4 % (Fig. 5D). The viability of the microspores did not change significantly during the culture process. Similar results were obtained with the Alexander staining (Fig. 5C) and the frequency of viability resulting from FDA was comparable with that of Alexander staining (Fig. 5E).Figure 5.


Reprogramming of cassava (Manihot esculenta) microspores towards sporophytic development.

Perera PI, Ordoñez CA, Dedicova B, Ortega PE - AoB Plants (2014)

Quantitative and qualitative demonstration of the microspore viability of tetrads of M. esculenta genotype TMS 60444. Fluorescein-diacetate-stained microspores of tetrads under (A) a light microscope and (B) a fluorescent microscope. (C) Microspores of tetrads stained with Alexander stain. Lane 1: tetrads with four viable microspores; Lane 2: with three viable microspores; Lane 3: with two viable microspores; Lane 4: with one viable microspore; Lane 5: without any viable microspores in the tetrads (scale bars = 20 µm). (D and E) Quantitative data of microspore viability in tetrads after sterilization (AS), 3 days after cold pretreatment (ACP), 3 days after culture (3DC) and 7 days after culture (7DC) as compared with fresh (F) tetrads with FDA (D) and Alexander staining (E).
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4061485&req=5

PLU022F5: Quantitative and qualitative demonstration of the microspore viability of tetrads of M. esculenta genotype TMS 60444. Fluorescein-diacetate-stained microspores of tetrads under (A) a light microscope and (B) a fluorescent microscope. (C) Microspores of tetrads stained with Alexander stain. Lane 1: tetrads with four viable microspores; Lane 2: with three viable microspores; Lane 3: with two viable microspores; Lane 4: with one viable microspore; Lane 5: without any viable microspores in the tetrads (scale bars = 20 µm). (D and E) Quantitative data of microspore viability in tetrads after sterilization (AS), 3 days after cold pretreatment (ACP), 3 days after culture (3DC) and 7 days after culture (7DC) as compared with fresh (F) tetrads with FDA (D) and Alexander staining (E).
Mentions: In order to find the reason for the low response in TMS, first the viability of the microspores in tetrads was evaluated by FDA and Alexander staining. The viability of the microspores in tetrads at the time of bud collection was compared with the viability at the different steps of culture protocol. Either all four microspores (Fig. 5, Lane 1) or three (Fig. 5, Lane 2), two (Fig. 5, Lane 3) or one microspore (Fig. 5, Lane 4) were viable in the tetrads when tested with FDA (Fig. 5A–C) in frequencies of 51.3, 25.2, 13.1 and 6 % in the freshly collected buds, whereas the frequency of tetrads with all four non-viable microspores (Fig. 5, Lane 5) was 4.4 % (Fig. 5D). The viability of the microspores did not change significantly during the culture process. Similar results were obtained with the Alexander staining (Fig. 5C) and the frequency of viability resulting from FDA was comparable with that of Alexander staining (Fig. 5E).Figure 5.

Bottom Line: In two cassava genotypes, different developmental stages of microspores were found to initiate sporophytic cell divisions, that is, with tetrads of TMS 60444 and with mid or late uni-nucleate microspores of SM 1219-9.A low frequency of the reprogramming and the presence of non-responsive microspores among the responsive ones in tetrads were found to be related to the viability and exine formation of the microspores.This paves the way for the development of an efficient technique for the production of homozygous lines in cassava.

View Article: PubMed Central - PubMed

Affiliation: Agrobiodiversity Research Area, International Center for Tropical Agriculture, A.A. 6713, Cali, Colombia Present address: Department of Horticulture and Landscape Gardening, Faculty of Agriculture and Plantation Management, Wayamba University, Gonawila, Makandura, Sri Lanka prasanthi1970@yahoo.com.

No MeSH data available.


Related in: MedlinePlus