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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.


Expression pattern of all 10 unigenes from the forward (A) and reverse (B) SSH library by real-time PCR.Shoots were treated with 20 μM COR and water. Cambia-containing tissues were collected half an hour (0.5 h), one hour (1 h), two hours (2 h), four hours (4 h), eight hours (8 h), one day (1 d), two days (2 d) and three days (3 d) after treatments. The relative expression was normalized to the housekeeping genes of HbACTIN and HbRH8. The data were shown as averages ± SE. *, significant difference (P < 0.05); **, very significant difference (P < 0.01).
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pone.0132070.g004: Expression pattern of all 10 unigenes from the forward (A) and reverse (B) SSH library by real-time PCR.Shoots were treated with 20 μM COR and water. Cambia-containing tissues were collected half an hour (0.5 h), one hour (1 h), two hours (2 h), four hours (4 h), eight hours (8 h), one day (1 d), two days (2 d) and three days (3 d) after treatments. The relative expression was normalized to the housekeeping genes of HbACTIN and HbRH8. The data were shown as averages ± SE. *, significant difference (P < 0.05); **, very significant difference (P < 0.01).

Mentions: The expression pattern of 10 unigenes from the forward and reverse SSH library was shown in Fig 4A and 4B, respectively. The Figs were drawn by the software OriginLab 9.0 (OriginLab Corporation. MA. USA). The expression pattern of most of the unigenes from the forward SSH library showed a sharp contrast between the COR and water treatments (Fig 4A). The unigenes encoding calmodulin-binding transcription activator (CAMTA), ubiquitin 11-like (UL), calcium-dependent protein kinase 1 gene (CDPK1), N-acetyl-farnesylcysteine (AFC) and galactono-1,4-lactone dehydrogenase (GLDH) were rapidly up-regulated at 1 h (CAMTA, UL, GLDH) and 2 h (CDPK1, AFC), and thereafter, they were down-regulated and remained relatively stable upon COR treatment (Fig 4A). It was noted that the transcript level of all of the four unigenes (UL, AFC, GLDH and CDPK1) was still significantly higher upon COR treatment than that upon water treatment at the late stage (from 8 h to 1d or 3 d). Only at the late stage (from 8 h to 3 d) was the transcript level of the unigene encoding for polygalacturonase (PGA) upon COR treatment significantly higher than that upon water treatment. The reverse was the case for the unigene encoding for CAMTA. By contrast, the transcript level of unigenes encoding indole-3-acetic acid-induced protein (ARG7) and calmodulin binding protein (CABP) at the early stage (from 1 h to 4 h) and the unigene encoding auxin-responsive protein IAA19 (IAA19) from 0.5 h to 1 d was significantly lower upon COR treatment than that upon water treatment. However, the expression patterns of most of the unigenes from the reverse SSH library were roughly similar between COR and water treatments (Fig 4B). Nevertheless, compared with the water treatment, the transcript levels of the unigenes encoding fructose-bisphosphate aldolase (FBA), phosphoinositide 5-phosphatase (5TPase) and sugar transporter (STP) were significantly higher while those encoding phenylalanine ammonia-lyase 3 (PAL), metallothionein (MT2) and mitochondrial cytochrome b (COB) were significantly lower at most of the time intervals after COR treatment. The transcript levels of the unigene encoding translation initiation factor-like protein (EIF) were significantly lower at the early stage (from 1 h to 4 h) and higher at the late stage (from 1 d to 3 d) after COR treatment.


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)

Expression pattern of all 10 unigenes from the forward (A) and reverse (B) SSH library by real-time PCR.Shoots were treated with 20 μM COR and water. Cambia-containing tissues were collected half an hour (0.5 h), one hour (1 h), two hours (2 h), four hours (4 h), eight hours (8 h), one day (1 d), two days (2 d) and three days (3 d) after treatments. The relative expression was normalized to the housekeeping genes of HbACTIN and HbRH8. The data were shown as averages ± SE. *, significant difference (P < 0.05); **, very significant difference (P < 0.01).
© Copyright Policy
Related In: Results  -  Collection

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

pone.0132070.g004: Expression pattern of all 10 unigenes from the forward (A) and reverse (B) SSH library by real-time PCR.Shoots were treated with 20 μM COR and water. Cambia-containing tissues were collected half an hour (0.5 h), one hour (1 h), two hours (2 h), four hours (4 h), eight hours (8 h), one day (1 d), two days (2 d) and three days (3 d) after treatments. The relative expression was normalized to the housekeeping genes of HbACTIN and HbRH8. The data were shown as averages ± SE. *, significant difference (P < 0.05); **, very significant difference (P < 0.01).
Mentions: The expression pattern of 10 unigenes from the forward and reverse SSH library was shown in Fig 4A and 4B, respectively. The Figs were drawn by the software OriginLab 9.0 (OriginLab Corporation. MA. USA). The expression pattern of most of the unigenes from the forward SSH library showed a sharp contrast between the COR and water treatments (Fig 4A). The unigenes encoding calmodulin-binding transcription activator (CAMTA), ubiquitin 11-like (UL), calcium-dependent protein kinase 1 gene (CDPK1), N-acetyl-farnesylcysteine (AFC) and galactono-1,4-lactone dehydrogenase (GLDH) were rapidly up-regulated at 1 h (CAMTA, UL, GLDH) and 2 h (CDPK1, AFC), and thereafter, they were down-regulated and remained relatively stable upon COR treatment (Fig 4A). It was noted that the transcript level of all of the four unigenes (UL, AFC, GLDH and CDPK1) was still significantly higher upon COR treatment than that upon water treatment at the late stage (from 8 h to 1d or 3 d). Only at the late stage (from 8 h to 3 d) was the transcript level of the unigene encoding for polygalacturonase (PGA) upon COR treatment significantly higher than that upon water treatment. The reverse was the case for the unigene encoding for CAMTA. By contrast, the transcript level of unigenes encoding indole-3-acetic acid-induced protein (ARG7) and calmodulin binding protein (CABP) at the early stage (from 1 h to 4 h) and the unigene encoding auxin-responsive protein IAA19 (IAA19) from 0.5 h to 1 d was significantly lower upon COR treatment than that upon water treatment. However, the expression patterns of most of the unigenes from the reverse SSH library were roughly similar between COR and water treatments (Fig 4B). Nevertheless, compared with the water treatment, the transcript levels of the unigenes encoding fructose-bisphosphate aldolase (FBA), phosphoinositide 5-phosphatase (5TPase) and sugar transporter (STP) were significantly higher while those encoding phenylalanine ammonia-lyase 3 (PAL), metallothionein (MT2) and mitochondrial cytochrome b (COB) were significantly lower at most of the time intervals after COR treatment. The transcript levels of the unigene encoding translation initiation factor-like protein (EIF) were significantly lower at the early stage (from 1 h to 4 h) and higher at the late stage (from 1 d to 3 d) after COR treatment.

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.