Limits...
Analysis of Differentially Expressed Genes Associated with Coronatine-Induced Laticifer Differentiation in the Rubber Tree by Subtractive Hybridization Suppression.

Zhang SX, Wu SH, Chen YY, Tian WM - PLoS ONE (2015)

Bottom Line: Pharmacological data show that jasmonic acid and its precursor linolenic acid are effective in inducing secondary laticifer differentiation in epicormic shoots of the rubber tree.The unigenes were associated with the stress/defense response, plant hormone signal transduction and structure development.It is suggested that Ca2+ signal transduction and redox seem to be involved in differentiation, while PGA and EIF are associated with the division of cambium initials for COR-induced secondary laticifer differentiation in the rubber tree.

View Article: PubMed Central - PubMed

Affiliation: College of Horticulture, Hainan University, Haikou, Hainan, 570228, China; Key Laboratory of Biology and Genetic Resources of Rubber Tree, Ministry of Agriculture, State Key Laboratory Incubation Base for Cultivation and Physiology of Tropical Crops, Rubber Research Institute, CATAS, Danzhou, Hainan, 571737, China.

ABSTRACT
The secondary laticifer in the secondary phloem is differentiated from the vascular cambia of the rubber tree (Hevea brasiliensis Muell. Arg.). The number of secondary laticifers is closely related to the rubber yield potential of Hevea. Pharmacological data show that jasmonic acid and its precursor linolenic acid are effective in inducing secondary laticifer differentiation in epicormic shoots of the rubber tree. In the present study, an experimental system of coronatine-induced laticifer differentiation was developed to perform SSH identification of genes with differential expression. A total of 528 positive clones were obtained by blue-white screening, of which 248 clones came from the forward SSH library while 280 clones came from the reverse SSH library. Approximately 215 of the 248 clones and 171 of the 280 clones contained cDNA inserts by colony PCR screening. A total of 286 of the 386 ESTs were detected to be differentially expressed by reverse northern blot and sequenced. Approximately 147 unigenes with an average length of 497 bp from the forward and 109 unigenes with an average length of 514 bp from the reverse SSH libraries were assembled and annotated. The unigenes were associated with the stress/defense response, plant hormone signal transduction and structure development. It is suggested that Ca2+ signal transduction and redox seem to be involved in differentiation, while PGA and EIF are associated with the division of cambium initials for COR-induced secondary laticifer differentiation in the rubber tree.

No MeSH data available.


Reverse northern blot analysis of differentially expressed ESTs in the forward and reverse SSH libraries.Panel A, B and C, hybridization with unsubtracted cDNA probes from samples upon COR treatment for 1 day (A), 2 days (B) and 3 days (C). Panels D, E and F, hybridization with unsubtracted cDNA probes from samples upon water treatment for 1 day (D), 2 days (E) and 3 days (F). a to k, ESTs from the forward SSH library. i to t, ESTs from the reverse SSH library. The strong and corresponding weak hybridization signals were indicated with solid rings and dotted rings, respectively.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4493031&req=5

pone.0132070.g002: Reverse northern blot analysis of differentially expressed ESTs in the forward and reverse SSH libraries.Panel A, B and C, hybridization with unsubtracted cDNA probes from samples upon COR treatment for 1 day (A), 2 days (B) and 3 days (C). Panels D, E and F, hybridization with unsubtracted cDNA probes from samples upon water treatment for 1 day (D), 2 days (E) and 3 days (F). a to k, ESTs from the forward SSH library. i to t, ESTs from the reverse SSH library. The strong and corresponding weak hybridization signals were indicated with solid rings and dotted rings, respectively.

Mentions: A total of 528 positive clones were obtained by blue-white screening, of which 248 clones came from the forward SSH library, while 280 clones came from the reverse SSH library. Approximately 215 of the 248 clones and 171 of the 280 clones contained cDNA inserts according to colony PCR screening. The differential expression of all the ESTs from the 386 positive clones (215 and 171 clones) was analyzed by reverse northern blot. The sheet containing 215 ESTs (Fig 2, line a-k) and 171 ESTs (Fig 2, line j-t) was hybridized with labeled cDNA probes from samples treated with COR (Fig 2A–2C) and water (Fig 2D–2F) for 1 day (Fig 2A and 2D), 2 days (Fig 2B and 2E) and 3 days (Fig 2C and 2F). A difference in dot color of the duplicates between hybridization with COR and water treatment at the same time interval was observed. A few of the differentially expressed spots were marked with real rings for strong dots and dotted rings for the corresponding weak ones (Fig 2).


Analysis of Differentially Expressed Genes Associated with Coronatine-Induced Laticifer Differentiation in the Rubber Tree by Subtractive Hybridization Suppression.

Zhang SX, Wu SH, Chen YY, Tian WM - PLoS ONE (2015)

Reverse northern blot analysis of differentially expressed ESTs in the forward and reverse SSH libraries.Panel A, B and C, hybridization with unsubtracted cDNA probes from samples upon COR treatment for 1 day (A), 2 days (B) and 3 days (C). Panels D, E and F, hybridization with unsubtracted cDNA probes from samples upon water treatment for 1 day (D), 2 days (E) and 3 days (F). a to k, ESTs from the forward SSH library. i to t, ESTs from the reverse SSH library. The strong and corresponding weak hybridization signals were indicated with solid rings and dotted rings, respectively.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4493031&req=5

pone.0132070.g002: Reverse northern blot analysis of differentially expressed ESTs in the forward and reverse SSH libraries.Panel A, B and C, hybridization with unsubtracted cDNA probes from samples upon COR treatment for 1 day (A), 2 days (B) and 3 days (C). Panels D, E and F, hybridization with unsubtracted cDNA probes from samples upon water treatment for 1 day (D), 2 days (E) and 3 days (F). a to k, ESTs from the forward SSH library. i to t, ESTs from the reverse SSH library. The strong and corresponding weak hybridization signals were indicated with solid rings and dotted rings, respectively.
Mentions: A total of 528 positive clones were obtained by blue-white screening, of which 248 clones came from the forward SSH library, while 280 clones came from the reverse SSH library. Approximately 215 of the 248 clones and 171 of the 280 clones contained cDNA inserts according to colony PCR screening. The differential expression of all the ESTs from the 386 positive clones (215 and 171 clones) was analyzed by reverse northern blot. The sheet containing 215 ESTs (Fig 2, line a-k) and 171 ESTs (Fig 2, line j-t) was hybridized with labeled cDNA probes from samples treated with COR (Fig 2A–2C) and water (Fig 2D–2F) for 1 day (Fig 2A and 2D), 2 days (Fig 2B and 2E) and 3 days (Fig 2C and 2F). A difference in dot color of the duplicates between hybridization with COR and water treatment at the same time interval was observed. A few of the differentially expressed spots were marked with real rings for strong dots and dotted rings for the corresponding weak ones (Fig 2).

Bottom Line: Pharmacological data show that jasmonic acid and its precursor linolenic acid are effective in inducing secondary laticifer differentiation in epicormic shoots of the rubber tree.The unigenes were associated with the stress/defense response, plant hormone signal transduction and structure development.It is suggested that Ca2+ signal transduction and redox seem to be involved in differentiation, while PGA and EIF are associated with the division of cambium initials for COR-induced secondary laticifer differentiation in the rubber tree.

View Article: PubMed Central - PubMed

Affiliation: College of Horticulture, Hainan University, Haikou, Hainan, 570228, China; Key Laboratory of Biology and Genetic Resources of Rubber Tree, Ministry of Agriculture, State Key Laboratory Incubation Base for Cultivation and Physiology of Tropical Crops, Rubber Research Institute, CATAS, Danzhou, Hainan, 571737, China.

ABSTRACT
The secondary laticifer in the secondary phloem is differentiated from the vascular cambia of the rubber tree (Hevea brasiliensis Muell. Arg.). The number of secondary laticifers is closely related to the rubber yield potential of Hevea. Pharmacological data show that jasmonic acid and its precursor linolenic acid are effective in inducing secondary laticifer differentiation in epicormic shoots of the rubber tree. In the present study, an experimental system of coronatine-induced laticifer differentiation was developed to perform SSH identification of genes with differential expression. A total of 528 positive clones were obtained by blue-white screening, of which 248 clones came from the forward SSH library while 280 clones came from the reverse SSH library. Approximately 215 of the 248 clones and 171 of the 280 clones contained cDNA inserts by colony PCR screening. A total of 286 of the 386 ESTs were detected to be differentially expressed by reverse northern blot and sequenced. Approximately 147 unigenes with an average length of 497 bp from the forward and 109 unigenes with an average length of 514 bp from the reverse SSH libraries were assembled and annotated. The unigenes were associated with the stress/defense response, plant hormone signal transduction and structure development. It is suggested that Ca2+ signal transduction and redox seem to be involved in differentiation, while PGA and EIF are associated with the division of cambium initials for COR-induced secondary laticifer differentiation in the rubber tree.

No MeSH data available.