Micro-scale chlorophyll analysis and developmental expression of a cytokinin oxidase/dehydrogenase gene during leaf development and senescence

Evans, Thomas G.; Song, Jiancheng; Jameson, Paula E.

doi:10.1007/s10725-011-9627-5

Cited by 11 publications

(8 citation statements)

References 22 publications

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“…Chlorophyll from three sub-samples of the pooled leaves for each developmental stage was extracted into DMF and analysed using a Nanodrop spectrophotometer as described in Evans et al [64]. The total chlorophyll content was calculated using the equation of Wellburn [65]: Chl total = 7·12 A 664 + 18·12 A 647.…”

Section: Methodsmentioning

confidence: 99%

Expression of Genes Related to Sugar and Amino Acid Transport and Cytokinin Metabolism during Leaf Development and Senescence in Pisum sativum L.

Ninan

Grant

Song

et al. 2019

Plants

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Gene editing is becoming the plant breeding tool of choice, but prior to targeting a gene for editing, a knowledge of the gene family members (GFMs) controlling yield is required in the specific crop plant. Critical to yield are components from senescing leaves. We targeted genes controlling senescence in Pisum sativum and the release and transport of carbohydrates and amino acids from the source leaves to the pods and seeds. The expression of GFMs for cytokinin biosynthesis (IPT) and destruction (CKX), sucrose transporters (SUT), Sugar Will Eventually be Exported Transporters (SWEET), amino acid permeases (AAP), and cell wall invertases, was determined using RT-qPCR. GFMs were differentially expressed in leaves of different ages. The expression of many gene family members was lower in the expanding sink leaves compared with the senescing leaves, with the exception of two PsAAP GFMs and PsCKX5, which were highly expressed. Expression of specific PsSWEETs, SUTs, and AAPs increased in the mature and/or senescing leaves. Expression of PsIPTs was least in the mature source leaves, but as strong in the senescing leaves as in the young source leaves. PsCKX7 was expressed in source and senescing leaves. We discuss the potential impact of the targeted reduction of specific PsCKX GFMs on source-sink relationships.

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Section: Methodsmentioning

confidence: 99%

Expression of Genes Related to Sugar and Amino Acid Transport and Cytokinin Metabolism during Leaf Development and Senescence in Pisum sativum L.

Ninan

Grant

Song

et al. 2019

Plants

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“…The chlorophyll content of cotyledon samples was measured from 4 hpi to 35 dpi using a Nanodrop spectrophotometer (Evans et al, 2012). The total chlorophyll content was calculated using the equation of Wellburn (1994): Chl total ¼ 7Á12A 664 þ 18Á12A 647…”

Section: Chlorophyll Estimationmentioning

confidence: 99%

Infection byRhodococcus fasciansmaintains cotyledons as a sink tissue for the pathogen

Dhandapani

Song

Novák

et al. 2016

Ann Bot

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Background and Aims Pisum sativum L. (pea) seed is a source of carbohydrate and protein for the developing plant. By studying pea seeds inoculated by the cytokinin-producing bacterium, Rhodococcus fascians, we sought to determine the impact of both an epiphytic (avirulent) strain and a pathogenic strain on source-sink activity within the cotyledons during and following germination.Methods Bacterial spread was monitored microscopically, and real-time reverse transcription-quantitative PCR was used to determine the expression of cytokinin biosynthesis, degradation and response regulator gene family members, along with expression of family members of SWEET, SUT, CWINV and AAP genes -gene families identified initially in pea by transcriptomic analysis. The endogenous cytokinin content was also determined.Key Results The cotyledons infected by the virulent strain remained intact and turned green, while multiple shoots were formed and root growth was reduced. The epiphytic strain had no such marked impact. Isopentenyl adenine was elevated in the cotyledons infected by the virulent strain. Strong expression of RfIPT, RfLOG and RfCKX was detected in the cotyledons infected by the virulent strain throughout the experiment, with elevated expression also observed for PsSWEET, PsSUT and PsINV gene family members. The epiphytic strain had some impact on the expression of these genes, especially at the later stages of reserve mobilization from the cotyledons.Conclusions The pathogenic strain retained the cotyledons as a sink tissue for the pathogen rather than the cotyledon converting completely to a source tissue for the germinating plant. We suggest that the interaction of cytokinins, CWINVs and SWEETs may lead to the loss of apical dominance and the appearance of multiple shoots.

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“…are responsible for marked heterogeneity among individual plants. The main problem with the methods cited above, and with more classical approaches like gas exchange measurements [ 6 ], is that they do not detect small differences in physiological traits between successive leaf ranks throughout the canopy.…”

Section: Introductionmentioning

confidence: 99%

A mobile NMR lab for leaf phenotyping in the field

et al. 2017

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BackgroundLow field NMR has been used to investigate water status in various plant tissues. In plants grown in controlled conditions, the method was shown to be able to monitor leaf development as it could detect slight variations in senescence associated with structural modifications in leaf tissues. The aim of the present study was to demonstrate the potential of NMR to provide robust indicators of the leaf development stage in plants grown in the field, where leaves may develop less evenly due to environmental fluctuations. The study was largely motivated by the need to extend phenotyping investigations from laboratory experiments to plants in their natural environment.MethodsThe mobile NMR laboratory was developed, enabling characterization of oilseed rape leaves throughout the canopy without uprooting the plant. The measurements made on the leaves of plants grown and analyzed in the field were compared to the measurements on plants grown in controlled conditions and analyzed in the laboratory.ResultsThe approach demonstrated the potential of the method to assess the physiological status of leaves of plants in their natural environment. Comparing changes in the patterns of NMR signal evolution in plants grown under well-controlled laboratory conditions and in plants grown in the field shows that NMR is an appropriate method to detect structural modifications in leaf tissues during senescence progress despite plant heterogeneity in natural conditions. Moreover, the specific effects of the environmental factors on the structural modifications were revealed.ConclusionThe present study is an important step toward the selection of genotypes with high tolerance to water or nitrogen depletion that will be enabled by further field applications of the method.

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Micro-scale chlorophyll analysis and developmental expression of a cytokinin oxidase/dehydrogenase gene during leaf development and senescence

Cited by 11 publications

References 22 publications

Expression of Genes Related to Sugar and Amino Acid Transport and Cytokinin Metabolism during Leaf Development and Senescence in Pisum sativum L.

Expression of Genes Related to Sugar and Amino Acid Transport and Cytokinin Metabolism during Leaf Development and Senescence in Pisum sativum L.

Infection byRhodococcus fasciansmaintains cotyledons as a sink tissue for the pathogen

A mobile NMR lab for leaf phenotyping in the field

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