Abstract
【Objective】 The objective of this study is to investigate the relationship between nitrate reductase(NR) and growth and regeneration of mulberry. A full length cDNA named MaNR was cloned from mulberry cultivar Guiyou No.62 and the sequence characteristics of MaNR were analyzed. Furthermore, the expression mode and influence factors during regeneration of mulberry hypocotyl regeneration in vitro was studied.【Method】 Specific primer sequences were designed according to the annotated gene scaffold 570 released in Morus notabilis genome database http://morus.swu.edu.cn/morusdb. Total RNA and DNA were extracted from mulberry leaf treated by KNO3, using RNAiso Plus(TaKaRa) and a modified CTAB method. cDNA and DNA were used as templates, RT-PCR was used to amplify MaNRcDNA sequence from RNA pools generated from KNO3-treated mulberry leaves. Bioinformatical tools were employed to analyze its deduced aimino acid sequence. Mulberry hypocotyls used as explants were inoculated in different nitrogen sources and growth regulating substances of the medium under aseptic conditions. Real-time PCR was used to investigate its relative expression level during hypocotyls regeneration in vitro and effects of different nitrogen sources and growth regulating substances. 【Result】 The obtained cDNA sequence of NR was 2 730 bp in length, encoding a polypeptide of 909 amino acid residues consisting of five classic conserved domain, protein molecular weight was 102.84 kD and the isoelectric point was 6.76. It showed that the sequence of NR has 95% identity compared with Morus notabilis and 78% indentity with other Rosaceae trees by NCBI BLAST. The obtained DNA sequence of NR was 5 142 bp including five exons and four introns. Cluster analysis showed that monocotyledons were clustered to one group, Mulberry and Rosaceae were clustered to one group, and the GenBank number was KF992020.1 and KF992021.1 respectively. Quantitative RT-PCR showed that this gene was highly expressed in roots, low expressed in stems. Glutamic acid significantly raised the expression of MaNR in leaves and GA3 significantly raised the expression of MaNR in stems. Hypocotyls could not be induced callus and multiple shoots in the single NH4+-N media but could be in the single NO3--N media. These nitrogen sources played a less part in subculture of shoot clumps. The shoots could grow in the single NH4+-N media and in the single NO3--N media. Real-time PCR exhibited that MaNR was highly abundant in cytoledons and hypocotyls than radicals. The relative expression level increased during hypocotyls regeneration in vitro and reduced to stable after shoot clumps formed. As time went on, the relative expression level of MaNR was reduced in shoot clumps with single NO3--N media or single NH4+-N. Glutamic acid showed a little inhibition of NR in the shoots subculture. 【Conclusion】 The full length of MaNR was cloned from mulberry trees. NO3--N was necessary for hypocotyls regeneration in vitro and the expression of NR affected by mulberry cell dedifferentiation and differentiation.
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Claims 20
Claim · model-written summary
The full-length MaNR cDNA was cloned from mulberry cultivar Guiyou No.62.
MorusOrganisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“of Morus L.”
e1“A full length cDNA named MaNR was cloned from mulberry cultivar Guiyou No.62”
Claim · model-written summary
The MaNR cDNA is 2 730 bp long and encodes a polypeptide of 909 amino acid residues.
- Gene
- NRe2
- Outcome
- cDNA sequencee2
- Value
- 2 730 bpe2
- Qualifier
- encoding a polypeptide of 909 amino acid residuese2
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“of Morus L.”
e2“The obtained cDNA sequence of NR was 2 730 bp in length, encoding a polypeptide of 909 amino acid residues”
Claim · model-written summary
The MaNR protein has a molecular weight of 102.84 kD.
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“of Morus L.”
e3“protein molecular weight was 102.84 kD and the isoelectric point was 6.76”
Claim · model-written summary
The MaNR protein has an isoelectric point of 6.76.
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“of Morus L.”
e3“protein molecular weight was 102.84 kD and the isoelectric point was 6.76”
Claim · model-written summary
The NR sequence is 95% identical to Morus notabilis.
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“of Morus L.”
e4“It showed that the sequence of NR has 95% identity compared with Morus notabilis and 78% indentity with other Rosaceae trees”
Claim · model-written summary
The NR sequence is 78% identical to other Rosaceae trees.
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“of Morus L.”
e4“It showed that the sequence of NR has 95% identity compared with Morus notabilis and 78% indentity with other Rosaceae trees”
Claim · model-written summary
The DNA sequence of NR is 5 142 bp with five exons and four introns.
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“of Morus L.”
e5“The obtained DNA sequence of NR was 5 142 bp including five exons and four introns”
Claim · model-written summary
Cluster analysis grouped monocotyledons into one group and Mulberry and Rosaceae into another group.
- Outcome
- Cluster analysise6
- Qualifier
- monocotyledons were clustered to one group, Mulberry and Rosaceae were clustered to one groupe6
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“of Morus L.”
e6“Cluster analysis showed that monocotyledons were clustered to one group, Mulberry and Rosaceae were clustered to one group”
Claim · model-written summary
MaNR was highly expressed in roots and expressed at a low level in stems.
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“of Morus L.”
e7“Quantitative RT-PCR showed that this gene was highly expressed in roots, low expressed in stems”
Claim · model-written summary
Glutamic acid significantly raised MaNR expression in leaves.
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“of Morus L.”
e1“Glutamic acid significantly raised the expression of MaNR in leaves and GA3 significantly raised the expression of MaNR in stems”
Claim · model-written summary
GA3 significantly raised MaNR expression in stems.
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“of Morus L.”
e1“Glutamic acid significantly raised the expression of MaNR in leaves and GA3 significantly raised the expression of MaNR in stems”
Claim · model-written summary
Hypocotyls did not form callus or multiple shoots on single NH4+-N media but did on single NO3--N media.
- Organ / stage
- Hypocotylse2
- Treatment
- single NH4+-N mediae2
- Outcome
- callus and multiple shootse2
- Comparator
- single NO3--N mediae2
Not reportedDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“of Morus L.”
e2“Hypocotyls could not be induced callus and multiple shoots in the single NH4+-N media but could be in the single NO3--N media”
Claim · model-written summary
Shoots could grow on single NH4+-N media.
Not reportedComparatorDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“of Morus L.”
e2“The shoots could grow in the single NH4+-N media and in the single NO3--N media”
Claim · model-written summary
Shoots could grow on single NO3--N media.
Not reportedComparatorDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“of Morus L.”
e2“The shoots could grow in the single NH4+-N media and in the single NO3--N media”
Claim · model-written summary
The relative expression level of MaNR increased during hypocotyl regeneration in vitro.
- Organ / stage
- hypocotylse2
- Outcome
- relative expression levele2
- Direction
- increasede2
- Qualifier
- during hypocotyls regeneration in vitroe2
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“of Morus L.”
e2“The relative expression level increased during hypocotyls regeneration in vitro and reduced to stable after shoot clumps formed”
Claim · model-written summary
The relative expression level of MaNR was reduced to a stable level after shoot clumps formed, including in shoot clumps on single NO3--N or single NH4+-N media.
- Treatment
- single NO3--N media or single NH4+-Ne3
- Gene
- MaNRe3
- Outcome
- relative expression levele3
- Direction
- reducede3
- Qualifier
- after shoot clumps formede2
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“of Morus L.”
e2“The relative expression level increased during hypocotyls regeneration in vitro and reduced to stable after shoot clumps formed”
e3“As time went on, the relative expression level of MaNR was reduced in shoot clumps with single NO3--N media or single NH4+-N”
Claim · model-written summary
MaNR was more abundant in cotyledons and hypocotyls than in radicals.
- Organ / stage
- cytoledons and hypocotylse2
- Gene
- MaNRe2
- Outcome
- MaNRe2
- Direction
- highly abundante2
- Comparator
- radicalse2
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“of Morus L.”
e2“Real-time PCR exhibited that MaNR was highly abundant in cytoledons and hypocotyls than radicals”
Claim · model-written summary
Nitrogen sources played a smaller part in the subculture of shoot clumps.
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“of Morus L.”
e2“These nitrogen sources played a less part in subculture of shoot clumps”
Claim · model-written summary
Glutamic acid showed a little inhibition of NR in shoot subculture.
- Organ / stage
- shootse2
- Treatment
- Glutamic acide2
- Outcome
- inhibition of NRe2
- Direction
- a little inhibitione2
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“of Morus L.”
e2“Glutamic acid showed a little inhibition of NR in the shoots subculture”
Claim · model-written summary
The authors interpret NR expression as affected by mulberry cell dedifferentiation and differentiation.
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“of Morus L.”
e2“the expression of NR affected by mulberry cell dedifferentiation and differentiation”
The same questions in other papers
in vitro · growth · Morus (mulberry trees)Positive and negative results
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- positive
- positive
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See all 10 papers →Claims are grouped by organism, treatment and what was measured or targeted. A paper counts as positive when any of its claims in the group is positive.