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Daily changes in the content of phytohormones accumulating in the hypocotyls of young sunflower seedlings that grow under a 14/10-h light-to-dark (LD) photoperiod were followed using LC–MS. The objective of the work was to investigate whether the presence of light and light transitions lead to visible changes visible in phytohormone levels on the sixth day after the onset of germination. The LC–MS technique used allowed the simultaneous detection of more than one hundred individual phytohormone derivatives, of which the daily accumulation patterns were determined for nearly fifty members. The daily changes in phytohormone levels followed specific patterns for each phytohormone. There were differences between the daytime and nighttime levels, most likely reflecting the effects of light on phytohormone metabolism. A significant difference was found between cytokinins (CKs) and all other phytohormones, so that CK group of phytohormones may be considered to have a separate role in hypocotyl elongation. Prolonged daytime (postponed dusk) resulted in a rapid disruption of rhythmic hypocotyl elongation and it triggered an acute light stress response that was evident in increased levels of a number of important phytohormones from all groups except CKs. However, this light stress, termed ARELD (Acute Response to Extended Light Duration), was not present in control plants grown under continuous light conditions. The data suggest that sunflower has means to adapt to the potentially stressful conditions of continuous light (LL). The qPCR-RT study of the isoforms of circadian clock-associated genes HaLHY, HaTOC1, HaELF3, and HaPIF3 revealed diurnal rhythms under both LD and LL conditions in which the peaks were synchronized and shifted toward the subjective dawn. [ABSTRACT FROM AUTHOR]

Light and gravity coordinately regulate the directional growth of plants. Arabidopsis Gravitropic in the Light 1 (GIL1) inhibits the negative gravitropism of hypocotyls in red and far‐red light, but the underlying molecular mechanisms remain elusive. Our study found that GIL1 is a plasma membrane‐localized protein. In endodermal cells of the upper part of hypocotyls, GIL1 controls the negative gravitropism of hypocotyls. GIL1 directly interacts with PIN3 and inhibits the auxin transport activity of PIN3. Mutation of PIN3 suppresses the abnormal gravitropic response of gil1 mutant. The GIL1 protein is unstable in darkness but it is stabilized by red and far‐red light. Together, our data suggest that light‐stabilized GIL1 inhibits the negative gravitropism of hypocotyls by suppressing the activity of the auxin transporter PIN3, thereby enhancing the emergence of young seedlings from the soil. [ABSTRACT FROM AUTHOR]

Mangroves are one of the world's most unique tropical coastal ecosystems. They are a rich repository of biological wealth, including specially adapted flora and fauna. The microbiome component of this ecosystem is a fascinating world that is yet to be fully explored for its functional and ecological interrelationships with its hosts. The mangrove ecosystem is a hidden treasure of microbial diversity, without which mangrove biology is incomplete. In the present study, the isolation of a cellulase-producing, endophytic Bacillus sp. from the hypocotyl region of viviparous seedlings is described. This study urges us to look into the microbial diversity of mangrove propagules, by presenting a glimpse of a member of the endospheric microbiome of viviparous hypocotyls. [ABSTRACT FROM AUTHOR]

High temperature-induced hypocotyl elongation is a typical thermomorphogenesis trait that may significantly affect early seedling growth and subsequent crop yield. The ambient temperature and endogenous auxin are two critical factors that regulate hypocotyl growth. However, the mechanism of temperature and auxin integration in horticultural plants remains poorly understood. In this study, the roles of the basic helix-loop-helix transcription factor CsPIF4 in regulating auxin biosynthesis genes and the auxin content in the hypocotyl of cucumber (Cucumis sativus L.) seedlings under high temperature were investigated. qRT-PCR and in situ hybridization analysis revealed that expression of CsPIF4 was enhanced in the epidermis and vascular bundles in the hypocotyl of cucumber seedlings in response to high temperature. qRT-PCR and HPLC analysis showed that CsPIF4 positively regulated transcription of the auxin biosynthesis gene CsYUC8 and the auxin content in the hypocotyl under high temperature (35 ℃). The CRISPR/Cas9-mediated knockout of CsPIF4 resulted in a shorter hypocotyl compared with that of the wild type, together with decreased expression of CsYUC8 and lower auxin content in response to high temperature. Furthermore, biochemical assays showed that CsPIF4 could bind directly to the G-box motif of the CsYUC8 promoter and thereby activate CsYUC8 expression. These findings provide insight into the molecular mechanism of high temperature-mediated hypocotyl elongation in cucumber. [ABSTRACT FROM AUTHOR]

This paper presents the results obtained from the analysis of physiological and biochemical parameters in the radish, species Raphanus sativus L., variety Rond Ecarlate, which was grown in three different systems (in the field, in the solarium and in the climate chamber). Several parameters necessary for the evaluation of biomass, physiological processes and nutritional values were determined. Regarding the biometric parameters, it was noticed that in the field cultivation the plants formed 11-12 leaves, in the solarium average 12-13 leaves, and in the climatic chamber the analyzed plants formed 7-9 leaves, on an average. The total biomass of the plant was recorded with the following average values: in the field 70.05 grams, in the solarium 92.3 grams and in the climate chamber 32.9 grams. Regarding the circumference of the tuberous hypocotyl axis, the values are higher in plants grown in solarium (16.9 cm), with the smallest dimensions being recorded in plants grown in the climate chamber (7.2 cm). The highest average pulp firmness value was observed in plants grown in the solarium (0.27 kg force), and the lowest values were seen in plants grown in the climate chamber (0.14 kg force). Regarding the amount of water lost through perspiration by the leaves of radish plants, the highest average values were recorded at plants grown in the solarium, and the lowest at plants grown in the climatic chamber. The average soluble dry matter content was higher (over 4%) in plants grown in the climate chamber compared to other harvesting systems. The analysis of the average carbohydrate content shows that the total soluble carbohydrates were present only in the hypocotyl axis and glucose was found both in the hypocotyl axis of the studied plants and in the leaves. Vitamin C was present in variable amounts, both in the hypocotyl axis and in the leaves. The average amino acid content (expressed in amino nitrogen) was between 0.003% and 0.006%. The anthocyanin was present with highest values (54.5 mg) in field-grown plants, and catalase activity was present both in the hypocotyl axis and in the leaves and roots. Air humidity and temperature had a high influence on the development of plant biomass, on the content of dry matter and on the loss of water through transpiration. The biometric, physiological and biochemical parameters varied depending on the cultivation system. [ABSTRACT FROM AUTHOR]

SUMMARY: Rapid hypocotyl elongation allows buried seedlings to emerge, where light triggers de‐etiolation and inhibits hypocotyl growth mainly by photoreceptors. Phosphorylation/dephosphorylation events regulate many aspects of plant development. Only recently we have begun to uncover the earliest phospho‐signaling responders to light. Here, we reported a large‐scale phosphoproteomic analysis and identified 20 proteins that changed their phosphorylation pattern following a 20 min light pulse compared to darkness. Microtubule‐associated proteins were highly overrepresented in this group. Among them, we studied CIP7 (COP1‐INTERACTING‐PROTEIN 7), which presented microtubule (MT) localization in contrast to the previous description. An isoform of CIP7 phosphorylated at Serine915 was detected in etiolated seedlings but was undetectable after a light pulse in the presence of photoreceptors, while CIP7 transcript expression decays with long light exposure. The short hypocotyl phenotype and rearrangement of MTs in etiolated cip7 mutants are complemented by CIP7‐YFP and the phospho‐mimetic CIP7S915D‐YFP, but not the phospho‐null CIP7S915A‐YFP suggesting that the phosphorylated S915CIP7 isoform promotes hypocotyl elongation through MT reorganization in darkness. Our evidence on Serine915 of CIP7 unveils phospho‐regulation of MT‐based processes during skotomorphogenic hypocotyl growth. Significance Statement: In our large‐scale phosphoproteome assay to elucidate the initial phospho‐signaling components triggered by a short light pulse, we found an enrichment of microtubule (MTs)‐associated proteins. Among these proteins, CIP7 (COP1‐INTERATING‐PROTEIN‐7) localizes to MTs and affects hypocotyl growth and MT orientation in dark hypocotyls. Serine915 of CIP7 is dephosphorylated by a brief light pulse. Phosphorylation status of Serine915 regulates this response. Our work establishes a link among skotomorphogenic development, phosphorylation dynamics and MTs‐based processes. [ABSTRACT FROM AUTHOR]

Ethylene plays essential roles in plant growth, development and stress responses. The ethylene signaling pathway and molecular mechanism have been studied extensively in Arabidopsis and rice but limited in peanuts. Here, we established a sand-culture method to screen pingyangmycin mutagenized peanut lines based on their specific response to ethylene (‘‘triple response‘‘). An ethylene-insensitive mutant, inhibition of peanut hypocotyl elongation 1 (iph1), was identified that showed reduced sensitivity to ethylene in both hypocotyl elongation and root growth. Through bulked segregant analysis sequencing, a major gene related to iph1, named AhIPH1, was preliminarily mapped at the chromosome Arahy.01, and further narrowed to a 450-kb genomic region through substitution mapping strategy. A total of 7014 genes were differentially expressed among the ACC treatment through RNA-seq analysis, of which only the Arahy.5BLU0Q gene in the candidate mapping interval was differentially expressed between WT and mutant iph1. Integrating sequence variations, functional annotation and transcriptome analysis revealed that a predicated gene, Arahy.5BLU0Q, encoding SNF1 protein kinase, may be the candidate gene for AhIPH1. This gene contained two single-nucleotide polymorphisms at promoter region and was more highly expressed in iph1 than WT. Our findings reveal a novel ethylene-responsive gene, which provides a theoretical foundation and new genetic resources for the mechanism of ethylene signaling in peanuts. [ABSTRACT FROM AUTHOR]

The current study aims of the current study is to establish cell suspension cultures of the medicinal plant, cumin Cuminum cyminum L. by the application of multiple drops and cross-section techniques. These methods were used in cultivating cell suspensions and in vitro plant regeneration. Leaf, stem, hypocotyl and root explants were cultured in Murashige and Skoog (MS) medium containing different concentrations (0.05, 0.5 and 1.0 mg.l-1) of naphthalene acetic acid (NAA) and (0.05, 0.1, 1.0 and 2.0 mg.l-1) benzyl adenine BA for callus production. The results indicated the high response of cumin, as the percentage of callus initiation was 100%.The plant regeneration percentage reached 91.6%. Moreover, a friable callus of hypocotyls was appropriate for initiation of cell suspension culture in MS medium with 0.5mg.l-1 NAA and 1.0 mg.l-1benzyl adenine (BA). The best density for primordial callus formation was 51.0 ×105 cells ml-1 in both multiple-drops and cross-sections embedding methods. Callus that had been produced from multi drops and sectors had the ability for shoot regeneration. This study clarified the efficiency of these techniques in establishing cell suspension culture and shoot regeneration, which could be promising sources for high production of active compounds in cumin plant.

The current study aims of the current study is to establish cell suspension cultures of the medicinal plant, cumin Cuminum cyminum L. by the application of multiple drops and cross-section techniques. These methods were used in cultivating cell suspensions and in vitro plant regeneration. Leaf, stem, hypocotyl and root explants were cultured in Murashige and Skoog (MS) medium containing different concentrations (0.05, 0.5 and 1.0 mg.l-1) of naphthalene acetic acid (NAA) and (0.05, 0.1, 1.0 and 2.0 mg.l-1) benzyl adenine BA for callus production. The results indicated the high response of cumin, as the percentage of callus initiation was 100%. The plant regeneration percentage reached 91.6%. Moreover, a friable callus of hypocotyls was appropriate for initiation of cell suspension culture in MS medium with 0.5mg.l-1 NAA and 1.0 mg.l-1benzyl adenine (BA). The best density for primordial callus formation was 51.0 ×105 cells ml-1 in both multiple-drops and cross-sections embedding methods. Callus that had been produced from multi drops and sectors had the ability for shoot regeneration. This study clarified the efficiency of these techniques in establishing cell suspension culture and shoot regeneration, which could be promising sources for high production of active compounds in cumin plant. [ABSTRACT FROM AUTHOR]

The difficult propagation of shrub and tree species and their extensive exposure to grazing threaten their abundance and lead to the necessity to find alternative means of propagation for these species. In vitro micropropagation techniques, viz., tissue culture, offer a promising tool for the rapid, cost-effective, and efficient propagation of different plant species. In the current study, a rapid and efficient in vitro multiplication protocol was developed for the micropropagation of Maerua crassifolia Forssk. Our results revealed that Murashige and Skoog (MS) medium with 7.5 µM of 6-benzylaminopurine (BA) and 1.0 µM of 1-naphthaleneacetic acid (NAA) led to the highest shoot formation (13.9 shoots per explant in 85.7% of the cultivated hypocotyls) among all other treatments. The best in vitro root formation was obtained on half-strength MS medium with 1.0 µM of indole 3-butyric acid (IBA) as 94.1% of the cultivated shoots formed 6.8 roots per microshoot on average. Ninety percent of the rooted plantlets were successfully acclimatized and are currently growing in the botanical garden of the Botany and Microbiology Department, King Saud University, Riyadh, Saudi Arabia. The genetic fidelity of the micropropagated plants was authenticated via flow cytometry. The results of the current study explained a simple, cost-effective, and efficient protocol for the micropropagation of the endangered M. crassifolia trees. [ABSTRACT FROM AUTHOR]

In vitro multiplication of okra is known to be delicate due to its recalcitrant nature. The current study was conducted to investigate the potential applications of thidiazuron (TDZ) on okra's in vitro regeneration. Cotyledonary leaves, hypocotyls and apices were taken from 15-day old in vitro grown seedlings, they were then inoculated on Gamborg (B5) medium containing different plant growth regulators. Maximum callus formation (100%) was obtained from cotyledonary leaves and hypocotyls on B5 without hormones also by adding 0.5 mgL-1 TDZ and 1 mgL-1 BAP for cotyledonary leaves followed by B5 containing 1 mgL-1 TDZ and 1 mgL-1 BAP with 95.83% of calli formed from hypocotyls. B5 supplemented with 1 mgL-1 of TDZ gave the highest rate (100%) of direct regeneration from apices followed by Gamborg medium without PGRs and B5 containing an equimolar concentration of TDZ and BAP with 66.66% of shoot formation. This regeneration system can be used to produce healthy plantlets with a low risk of genetic instability and somaclonal variants. [ABSTRACT FROM AUTHOR]

Copyright of Chinese Journal of Oil Crop Sciences is the property of Oil Crops Research Institute of Chinese Academy of Agricultural Sciences and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Environmental & Social Management Journal / Revista de Gestão Social e Ambiental is the property of Environmental & Social Management Journal and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Among the most common abiotic stresses that retard the development and growth of plants is salt stress. This study examined the impact of various salt levels (NaCl and Na2SO4) on the germination behaviour of sesame cultivars. Under various simulated salt concentrations (0.05M, 0.01M,0.15M,0.2M,0.25M,0.3M,0.35M,0.4M, and 0.5M), the degree of salinity tolerance of two sesame cultivars, viz., GT-10 and JTS-8 were assessed. The radicle, hypocotyl length, and fresh weight measurements were made separately after 96 and 144 hours. The results revealed that germination and seedling growth drastically decreased as the salt concentration increased. In comparison to GT-10, JTS-8 exhibits greater tolerance to different NaCl concentrations and greater sensitivity to varying Na2SO4 concentrations. [ABSTRACT FROM AUTHOR]

To establish a method for studying the organic acid environmental capacity of mangrove ecosystems, high-performance liquid chromatography was used to measure the organic acid detoxification agent; Using different cultivation methods to determine the toxicity threshold of organic acids on mangrove plants; Calculate the environmental capacity of organic acids by combining the toxicity threshold of organic acids with the volume of water in the study area. The results showed the range of toxicity thresholds of organic acids to 25.29–30 mg/L would have an inhibitory effect on the development of mangrove plant hypocotyls; The organic acid environmental capacity of Dongzhai harbor Mangrove Wetland Protection Area is 7.76 × 10^4 kg/d ~ 8.73 × 10^4 kg/d, while the estimated organic acid emissions from shrimp ponds around Dongzhai harbor are 7.06 × 10^3 kg/d ~ 7.83 × 10^3 kg/d. Therefore, the organic acid emissions from shrimp ponds around Dongzhai harbor are within the carrying range of the mangrove wetland ecosystem in Dongzhai harbor. • A research method on the environmental capacity of organic acids in mangrove ecosystems was established, based on the determination of RT-HPLC methods for the simultaneous detection of a variety of organic acids, and the testing of commercially available organic acid detoxifiers to clarify the types and concentrations of organic acids used in the experiments. • Concentration ranges of organic acids discharged into mangrove wetland ecosystems obtained from laboratory experiments modelling the degradation kinetics of organic acid detoxifiers in use. • Determine the toxicity thresholds of organic acids to mangrove plants using both hydroponic and soil cultivation methods; calculate the environmental capacity of mangrove wetland reserves for organic acids by combining the toxicity thresholds of organic acids with the volume of water in the research area. [ABSTRACT FROM AUTHOR]

Paubrasilia echinata (brazilwood) is an endangered native tree from the Brazilian Atlantic Forest whose seeds tolerate maturation drying, but, unlike classic orthodox seeds, they quickly lose viability after shedding. This work analyzed the biochemical and ultrastructural changes during the maturation of brazilwood seeds, with particular attention to the cell walls and organization of the cellular components. The physiological seed maturity was accompanied by increased starch content and decreased soluble sugars. Arabinose increased considerably and was the predominant cell-wall sugar during maturation, suggesting a rise in arabinans that contribute to greater cell wall flexibility. This increase was consistent with the cell wall infolding observed in the hypocotyl axis and cotyledons during the maturation of brazilwood seeds. Ultrastructural analyses showed changes in the number and distribution of protein bodies and amyloplasts and the reorganization of lipid droplets into large drops or masses during seed desiccation. Our findings demonstrate that brazilwood seeds behave like other orthodox seeds during maturation, performing the cell wall and metabolic changes before the major decline in the seed water content. However, the high vacuolization and reorganization of lipid bodies observed at 65 DAA suggest that cell deterioration occurs to some extent at the end of the maturation period and could be responsible for reducing the longevity of the brazilwood dried seeds. [ABSTRACT FROM AUTHOR]

The difficult propagation of shrub and tree species and their extensive exposure to grazing threaten their abundance and lead to the necessity to find alternative means of propagation for these species. In vitro micropropagation techniques, viz., tissue culture, offer a promising tool for the rapid, cost-effective, and efficient propagation of different plant species. In the current study, a rapid and efficient in vitro multiplication protocol was developed for the micropropagation of Maerua crassifolia Forssk. Our results revealed that Murashige and Skoog (MS) medium with 7.5 µM of 6-benzylaminopurine (BA) and 1.0 µM of 1-naphthaleneacetic acid (NAA) led to the highest shoot formation (13.9 shoots per explant in 85.7% of the cultivated hypocotyls) among all other treatments. The best in vitro root formation was obtained on half-strength MS medium with 1.0 µM of indole 3-butyric acid (IBA) as 94.1% of the cultivated shoots formed 6.8 roots per microshoot on average. Ninety percent of the rooted plantlets were successfully acclimatized and are currently growing in the botanical garden of the Botany and Microbiology Department, King Saud University, Riyadh, Saudi Arabia. The genetic fidelity of the micropropagated plants was authenticated via flow cytometry. The results of the current study explained a simple, cost-effective, and efficient protocol for the micropropagation of the endangered M. crassifolia trees.

In this study, we used the modified CRISPR/Cas9 system to produce targeted point mutations in cauliflower. Acetolactate synthase (ALS) and Centromere-specific histone H3 variant (CENH3) genes were selected as the base-editing targets and hypocotyls of cauliflower were used as explants. For ALS gene, a C-to-T conversion in the Pro182 codon (CCT) can alter the encoded amino acid, likely resulting in herbicide resistance, and a C-to-T mutation in the Leu133 codon (CTT) in the CENH3 gene may produce a haploid inducer. Results indicated that the transformation efficiency was 1.8%-4.5% and the mutation efficiencies for the ALS and CENH3 genes were approximately 22% and 87%, respectively. The ALS mutant cauliflower showed strong herbicide resistance, with possible immediate implications for broadleaf weed control in cauliflower fields. [ABSTRACT FROM AUTHOR]

NHX5 and NHX6, Arabidopsis endosomal antiporters, play a vital role in facilitating ion and pH homeostasis in endosomal compartments. Studies have found that NHX5 and NHX6 are essential for protein trafficking, auxin homeostasis, and plant growth and development. Here, we report the role of NHX5 and NHX6 in brassinosteroid (BR) signaling. We found that hypocotyl growth was enhanced in nhx5 nhx6 under epibrassinolide (eBR) treatment. nhx5 nhx6 bri1 was insensitive to eBR treatment, indicating that NHX5 and NHX6 are downstream of the BRI1 receptor in BR signaling. Moreover, confocal observation with both hypocotyls and root tips showed that BRI1-YFP localization in the plasma membrane (PM) was reduced in nhx5 nhx6. Interestingly, brefeldin A (BFA) treatment showed that formation of the BFA bodies containing BRI1 and their disassembling were disrupted in nhx5 nhx6. Further genetic analysis showed that NHX5/NHX6 and SYP22 may act coordinately in BR signaling. NHX5 and NHX6 may regulate SYP22 function by modulating cellular K+ and pH homeostasis. Importantly, NHX5 and NHX6 colocalize and interact with SYP22, but do not interact with BRI1. In summary, our findings indicate that NHX5/NHX6/SYP22 complex is essential for the regulation of BRI1 recycling and PM localization. The H+-leak facilitated by NHX5 and NHX6 offers a means of controlling BR signaling in plants. [ABSTRACT FROM AUTHOR]

Microcystin-LR (MC-LR) is a harmful cyanotoxin that inhibits 1 and 2A serine-threonine protein phosphatases. This study examines the influence of MC-LR on chloroplast division and the underlying mechanisms and consequences in Arabidopsis. MC-LR increased the frequency of dividing chloroplasts in hypocotyls in a time range of 1–96 h. At short-term exposures to MC-LR, small-sized chloroplasts (longitudinal diameters ≤6 μm) were more sensitive to these stimulatory effects, while both small and large chloroplasts showed stimulations at long-term exposure. After 48 h, the cyanotoxin increased the frequency of small-sized chloroplasts, indicating the stimulation of division. MC-LR inhibited protein phosphatases in whole hypocotyls and isolated chloroplasts, while it did not induce oxidative stress. We show for the first time that total cellular phosphatases play important roles in chloroplast division and that particular chloroplast phosphatases may be involved in these processes. Interestingly, MC-LR has a protective effect on cyanobacterial division during methyl-viologen (MV) treatments in Synechococcus PCC6301. MC-LR production has harmful effects on ecosystems and it may have an ancient cell division regulatory role in stressed cyanobacterial cells, the evolutionary ancestors of chloroplasts. We propose that cytoplasmic (eukaryotic) factors also contribute to the relevant effects of MC-LR in plants. [Display omitted] • Microcystin-LR (MC-LR) is a harmful cyanotoxin, an inhibitor of protein phosphatases. • MC-LR alters Arabidopsis plastid division at environmentally relevant concentration. • Environmental implications of such subcellular alterations in plants are discussed. • MC-LR also affects cyanobacterial division even during oxidative stress. • MC-LR's evolutionary function may be the regulation of cyanobacterial division. [ABSTRACT FROM AUTHOR]

Winter rape (Brassica napus L.) is better than other edible oil crops in China, but poor cold resistance is the key factor restricting its development. Hypocotyl length was found closely related to cold tolerance. The correlation between hypocotyl length and semi-lethal low temperature was significant, and the highest correlation between hypocotyl length and LT50 of autumn sowing was 0.9557. When the hypocotyl were treated at low temperature, the cells were seriously damaged and formed cavity structures, with cell walls seriously damaged or merged into each other. The positive regulation gene of hypocotyl length in resistant line of VHTSG 10 was identified as HY5 (transcription factor HY5-like) by qPCR, and bZIP transcription factor was found to be its conserved domain. Fused gene by GFP and HY5 from VHTSG 10 was transient transferred into Nicotiana benthamiana cells. Corresponding to the 35S:GFP widely distributed in plasma membrane of leaf epidermis, the fusion protein 35S:HY5-GFP was mainly distributed in nucleus. Thus we regarded BnHY5 gene is a key gene related to cold tolerance and hypocotyl length in B. napus. [ABSTRACT FROM AUTHOR]

Mobilisation of seed storage reserves is important for seedling establishment in Arabidopsis. In this process, sucrose is synthesised from triacylglycerol via core metabolic processes. Mutants with defects in triacylglycerol-to-sucrose conversion display short etiolated seedlings. We found that whereas sucrose content in the indole-3-butyric acid response 10 (ibr10) mutant was significantly reduced, hypocotyl elongation in the dark was unaffected, questioning the role of IBR10 in this process. To dissect the metabolic complexity behind cell elongation, a quantitative-based phenotypic analysis combined with a multi-platform metabolomics approach was applied. We revealed that triacylglycerol and diacylglycerol breakdown were disrupted in ibr10, resulting in low sugar content and poor photosynthetic ability. Importantly, batch-learning self-organised map clustering revealed that threonine level was correlated with hypocotyl length. Consistently, exogenous threonine supply stimulated hypocotyl elongation, indicating that sucrose levels are not always correlated with etiolated seedling length, suggesting the contribution of amino acids in this process. [ABSTRACT FROM AUTHOR]

Mitochondria in plant cells usually contain less than a full copy of the mitochondrial DNA (mtDNA) genome. Here, we asked whether mitochondrial dynamics may allow individual mitochondria to 'collect' a full set of mtDNA-encoded gene products over time, by facilitating exchange between individuals akin to trade on a social network. We characterise the collective dynamics of mitochondria in Arabidopsis hypocotyl cells using a recent approach combining single-cell time-lapse microscopy, video analysis and network science. We use a quantitative model to predict the capacity for sharing genetic information and gene products through the networks of encounters between mitochondria. We find that biological encounter networks support the emergence of gene product sets over timemore readily than a range of other possible network structures. Using results from combinatorics, we identify the network statistics that determine this propensity, and discuss how features of mitochondrial dynamics observed in biology facilitate the collection of mtDNA-encoded gene products. [ABSTRACT FROM AUTHOR]

C8 alcohols, such as 1‐octen‐3‐ol and 3‐octanol, have been variously described as mushroom, musty or earthy and are common volatile compounds of soymilk. C8 alcohols were the major volatiles formed during soybean soaking, and about 95% of the total C8 alcohols were originated from the enzymatic hydrolysis of glycosides of alcohols in soybean hypocotyls. In addition to 1‐octen‐3‐yl β‐primeveroside, a new glycoside was purified from the methanol extracts of soybean hypocotyls and were confirmed as 3‐octanyl β‐primeverosides by nuclear magnetic resonance (NMR) and high‐resolution mass spectrometry (HRMS). The crude glycosidase extracted from soybean hypocotyls showed much higher hydrolysis activities toward C8 alcohol β‐primeverosides than that of almond β‐glycosidase. Besides, increasing nonenzymatic hydrolysis was observed when the purified β‐primeverosides were heated at pH values lower than 6. The nonenzymatic hydrolysis activation energies (Ea) were 30.41 kJ/mol and 38.08 kJ/mol for 1‐octen‐3‐yl β‐primeveroside and 3‐octanyl β‐primeveroside, respectively. In addition to C8 alcohols, nonenzymatic hydrolysis also resulted in the formation of 2,3‐octanedione, 3‐hydroxy‐2‐octanone and 3‐octanone. The nonenzymatic hydrolysis of β‐primeverosides in the soy milk was also studied under the same conditions as those in the model systems. The degree of hydrolysis was similar but the molar yields of the volatile components were lower than those obtained in the model system. Practical Application: The results indicated that a different strategy or technology is required to suppress the formation of C8 alcohols than that applied to off‐flavor compounds formed de novo during processing of soybean seeds. [ABSTRACT FROM AUTHOR]

Allelopathic effects of the donor plants are mostly regarded as harmful to target plants such as traditional crops. Traditional crops play pivotal role as staple food for rural communities and hence ensuring food security. Study of the allelopathic effects of Androstachys johnsonii Prain on the germination and growth of Zea mays L. and Vigna subterranea (L.) Verdc was conducted. Different plant leachates concentrations of 0.675%, 1.25%, 2.5% and 5% were prepared through serial dilutions. Germination bioassays were arranged in a complete randomized setup of three replicates for each treatment for seven days. Germination rates were calculated, radicle and hypocotyl lengths were also measured. Germination rates of Z. mays L. and V. subterranea (L.) Verdc. ranged from 50 - 100% and 18 - 76%, respectively. Significant differences of P< 0.005 were noticed from the germination rates of V. subterranea (L.) Verdc subjected to high concentrations of different extracts used. Statistically there was no significant differences between germination rates of Z. mays L. treated with all concentrations of leaf leachates against germination rates of the seeds treated with distilled water. Outstandingly, the germination rate and early growth of Z. mays L. were promoted by less concentrated extracts of leaf, root, stem bark and soil which then suggest future use of allelochemicals from Androstachys johnsonii Prain for promotion of maize plant productions. Based on the beneficial attributes of allelochemicals from Androstachys johnsonii on the germination and growth of Z. mays L., planting of Androstachys johnsonii Prain in maize fields is highly recommended. [ABSTRACT FROM AUTHOR]

The leading purpose of this work is the development of efficient culture conditions to induce calli from cabbage (Brassica oleracea var. capitata L.) under in vitro conditions. The mature seeds were surface sterilized with combinations of different concentrations of ethanol and NaOCl in different time durations and were germinated on MS basal medium. The results revealed that the best sterilization method of cabbage seeds was by using 70% ethanol for one minute, followed by 15 min in 2% (NaOCl). Seedlings were used as donor sources for hypocotyls, cotyledon leaves, true leaves, and shoot tip explants. These explants were cultured on different combinations of cytokinins (TDZ, BAP, Ad) and auxins (IAA, NAA, 2, 4-D) then implanted in Murashige and Skoog (MS) media. 4 weeks after culturing, a significant difference was found among the explants in response to plant hormones. The maximum percentage of callus induction (100%) was using the combinations of 1 BAP + 1 2, 4-D, 1 BAP + 1 NAA, and 1 BAP + 2 2,4-D mg. l-1 . In addition, explants responses varied and the hypocotyls showed a superior result (85.71 %) as compared to other explants. For callus fresh weight, the combination of 0.22 TDZ + 79.9 Ad mg. l-1 had a significant effect, causing the highest fresh weight (0.2745g), while control treatment gave the lowest mean of 0.0066 g. Data showed that cotyledon explants were significantly superior in giving highest callus fresh weight with the mean of 0.1723 g. On the other hand, hypocotyl explants gave the lowest mean, reaching 0.1542 g. [ABSTRACT FROM AUTHOR]

Peanut (Arachis hypogaea L.) seeds were germinated to investigate the effect of the fermentation period of oak tree sawdust on germination viability and seedling characteristics. Its germination rate, seedling weight, length, and total vigor index were assessed. The seeds were sown in oak tree sawdust fermented for 0, 30, 45, and 60 days. The germination rates of the seeds in fermented sawdust were significantly different. The seeds in the 45-day fermented sawdust produced the heaviest biomass weight (4.6 g) with the longest true leaf (1.7 cm) and hypocotyl (3.4 cm) resulting in the highest total vigor index (925.8). In contrast, seeds in 0-day fermented sawdust had the lowest total vigor index (18.3). Microbiome analysis showed that the microbial community in the sawdust changed as the fermentation progressed, indicating that the microbial community seems to affect seed germination physiology. Taken together, 45-day fermented sawdust is recommended for optimal peanut seed germination and seedling growth. [ABSTRACT FROM AUTHOR]

The hypocotyls of germinating seedlings elongate in a search for light to enable autotrophic sugar production. Upon exposure to light, photoreceptors that are activated by blue and red light halt elongation by preventing the degradation of the hypocotyl-elongation inhibitor HY5 and by inhibiting the activity of the elongation-promoting transcription factors PIFs. The question of how sugar affects hypocotyl elongation and which cell types stimulate and stop that elongation remains unresolved. We found that overexpression of a sugar sensor, Arabidopsis hexokinase 1 (HXK1), in guard cells promotes hypocotyl elongation under white and blue light through PIF4. Furthermore, expression of PIF4 in guard cells is sufficient to promote hypocotyl elongation in the light, while expression of HY5 in guard cells is sufficient to inhibit the elongation of the hy5 mutant and the elongation stimulated by HXK1. HY5 exits the guard cells and inhibits hypocotyl elongation, but is degraded in the dark. We also show that the inhibition of hypocotyl elongation by guard cells' HY5 involves auto-activation of HY5 expression in other tissues. It appears that guard cells are capable of coordinating hypocotyl elongation and that sugar and HXK1 have the opposite effect of light on hypocotyl elongation, converging at PIF4. Kelly et al. show that Arabidopsis hexokinase1 (HXK1) expressed in guard-cells is sufficient to drive hypocotyl elongation through increasing the activity of PIF4 and auxin level, and competing with the effects of HY5. This study provides insights into how light and sucrose antagonistically coordinate the effort to achieve the height necessary for efficient photosynthetic, autotrophic sugar production. [ABSTRACT FROM AUTHOR]

Fusarium graminearum is the most devastating pathogen of Fusarium head blight of cereals, stalk and ear of maize, and it has recently become a potential threat for soybean as maize-soybean strip relay intercropping is widely practiced in China. To elucidate the pathogenesis mechanism of F. graminearum on intercropped soybean which causes root rot, transcriptional profiling of F. graminearum at 12, 24, and 48 h post-inoculation (hpi) on soybean hypocotyl tissues was conducted. In total, 2313 differentially expressed genes (DEGs) of F. graminearum were annotated by both KEGG pathway and Gene Ontology (GO) analysis. Among them, 128 DEGs were commonly expressed at three inoculation time points while the maximum DEGs were induced at 24 hpi. In addition, DEGs were also rich in carbon metabolism, ribosome and peroxisome pathways which might contribute to carbon source utilization, sexual reproduction, virulence and survival of F. graminearum when infected on soybean. Hence, this study will provide some basis for the deep understanding the pathogenesis mechanism of F. graminearum on different hosts and its effective control in maize-soybean strip relay intercropping systems. [ABSTRACT FROM AUTHOR]

Trachyspermum ammi L. Sprague, commonly known as ajwain, carom or thymol seeds, is one of the oldest medicinal plants with numerous health benefits. The classical breeding process of medicinal plants is not only time consuming but also requires much efforts. Biotechnological techniques such as in vitro regeneration which increase the capacity of production of medicinal plants rely upon a plenty of components. Here, we studied regeneration of T. ammi by indirect somatic embryogenesis by employing MS and B5 media supplemented with a diverse concentration of plant growth regulators (PGRs), including 2,4-D, kinetin, NAA, IAA, and BAP with four explants (hypocotyl, epicotyl, root and node) and 3 ecotypes (Qom, Yazd, and Rafsanjan). The highest callus induction was obtained from hypocotyl explants on MS medium supplemented with 8.917 Îm/L 2,4-D and 2.32 Îm/L KIN and epicotyl explants on B5 medium supplemented with 1.074 Îm/L NAA and 0.89 Îm/L BAP. The highest indirect regeneration (83%) was observed in hypocotyl explants on MS medium supplemented with 0.93 Îm/L kinetin. To the best of our knowledge, the present study (2017) is the first report on indirect somatic embryogenesis from hypocotyl and epicotyl of the Iranian Trachyspermum ammi ecotypes that can be beneficial for genetic transformation and other plant biotechnology techniques. [ABSTRACT FROM AUTHOR]

In this study, calli of Medicago sativa L. cv. Elçi (alfalfa Elçi) were inoculated in cell suspension culture and analyzed for aggregate assay, cell viability test, total phenolic content assay, DPPH free radical scavenging activity and formononetin assay by means of High-Performance Liquid Chromatography (HPLC). Hypocotyl, cotyledon and apical meristem explants were taken from 15-day-old aseptic seedlings and germinated in MS medium. 10 g calli were grown for each explant and then transferred into cell suspension culture. The highest cell viability rate, which was 75%, and the highest DPPH free radical scavenging activity with 51.36% was measured in 1000 mL cell suspension culture, while the highest total phenolic content, i.e. 40.2 mg/g, was quantified in 250 mL cell suspension culture. In accordance with the findings of the study, the production of formononetin was higher in the calli derived from cell suspension cultures than in herb samples of M. sativa. Moreover, in 1000 mL cell suspension culture, 4.99 mg/g of formononetin concentration was quantified, which scored the highest. In large-scale cell suspension cultures of M. sativa, it was possible to increase the production of formononetin production. Hence, due to its medicinal significance, a method has been tested to obtain higher amounts of this compound. [ABSTRACT FROM AUTHOR]

In plants, alternative splicing (AS) is markedly induced in response to environmental stresses, but it is unclear why plants generate multiple transcripts under stress conditions. In this study, RNA-seq was performed to identify AS events in cucumber seedlings grown under different light intensities. We identified a novel transcript of the gibberellin (GA)-deactivating enzyme Gibberellin 2-beta-dioxygenase 8 (CsGA2ox8). Compared with canonical CsGA2ox8.1, the CsGA2ox8.2 isoform presented intron retention between the second and third exons. Functional analysis proved that the transcript of CsGA2ox8.1 but not CsGA2ox8.2 played a role in the deactivation of bioactive GAs. Moreover, expression analysis demonstrated that both transcripts were upregulated by increased light intensity, but the expression level of CsGA2ox8.1 increased slowly when the light intensity was >400 µmol·m−2·s−1 PPFD (photosynthetic photon flux density), while the CsGA2ox8.2 transcript levels increased rapidly when the light intensity was >200 µmol·m−2·s−1 PPFD. Our findings provide evidence that plants might finely tune their GA levels by buffering against the normal transcripts of CsGA2ox8 through AS. [ABSTRACT FROM AUTHOR]

The circadian clock coordinates an organism's growth, development and physiology with environmental factors. One illuminating example is the rhythmic growth of hypocotyls and cotyledons in Arabidopsis thaliana. Such daily oscillations in leaf position are often referred to as sleep movements or nyctinasty. Here, we report that plantlets of the liverwort Marchantia polymorpha show analogous rhythmic movements of thallus lobes, and that the circadian clock controls this rhythm, with auxin a likely output pathway affecting these movements. The mechanisms of this circadian clock are partly conserved as compared to angiosperms, with homologs to the core clock genes PRR, RVE and TOC1 forming a core transcriptional feedback loop also in M. polymorpha. [ABSTRACT FROM AUTHOR]

Copyright of Journal of Agriculture & Nature / Kahramanmaraş Sütçü İmam Üniversitesi Tarım & Doğa Dergisi is the property of Kahramanmaras Sutcu Imam Universitesi and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Agronomía Colombiana is the property of Universidad Nacional de Colombia and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Contaminated water with hexavalent chromium Cr(VI) is a serious environmental problem. This study aimed to evaluate the Cr(VI) removal by zero valent iron nanoparticles (nZVI) reduction process and the impact of Cr(VI), nZVI and combined treatment with nZVI and Cr(VI) on tomato growth performance. To evaluate the Cr(VI) toxic effect on germination capability, seeds were exposed to increasing Cr(VI) concentrations up to 1000 mg L−1. The inhibition of seed germination and the decrease of hypocotyl and root length started from Cr(VI) 5 mg L−1. Under treatment with Cr(VI) + nZVI 5 mg L−1, seed germination, hypocotyl and root length resulted significantly higher compared to Cr(VI) 5 mg L−1 treatment. The impact of only nZVI was investigated on chlorophyll and carotenoid in leaves; iron levels in leaves, roots, fruits and soil; carotenoid, fat-soluble vitamin and nicotianamine in mature fruits. A significant increase of leaf chlorophyll and carotenoids was observed after nZVI 5 mg L−1 treatment compared to controls. No significant variations were observed in carotenoids, fat-soluble vitamins and nicotianamine levels after treatment with nZVI 5 mg L−1 in mature fruits. For their ability to reduce Cr(VI) and to stimulate tomato growth, nZVI might to be considered as alternative for remediation purposes. [ABSTRACT FROM AUTHOR]

The aim of the research was to improve the genetic transformation and adaptation conditions of commercial winter rapeseed line Brassica napus L. with subsequent production of transgenic seeds. It was optimized the technique of Agrobacterium-mediated genetic transformation of winter rapeseed Bn1 line (Brassica napus L.) using hypocotyls of 6- day-old seedlings as explants. GUS activity histochemical analysis showed a positive reaction in calli cultures and regenerant plant leaves obtained after Agrobacterium tumefaciens GV3101 transformation, which carried pCB203 plasmid with gus and bar genes. The optimal transformation conditions were determined as follows: the bacterial suspension optical density at a 600 nm wavelength - OD = 0.5, 10 min of inoculation and 48 h of co-cultivation. Stable integration of marker genes was confirmed by both histochemical and PCR analysis. [ABSTRACT FROM AUTHOR]

Development of an efficient in vitro plant regeneration system plays an important role for applying of biotechnological approaches for crop improvement. Therefore, the effect of added of BAP in germination phase on organogenesis of cotyledon and hypocotyl explants of two melon lines was investigated. The seeds germination medium contained three different concentrations of BAP (0.5, 1.0 and 1.5 mg L-1). The rate of plant regeneration was found to depend on genotype, explant type and culture medium. The cotyledons were more effective as explants for organogenesis and subsequent plant elongation than hypocotyls. Combination of 1.0 mg L-1 of BAP in germination medium and 0.5 mg L-1 BAP + 0.5 mg L-1 IAA in next regeneration medium gives the better regeneration answer in two explant types of line 11/9, while in line AGY the most effective was the combination of 1.5 mg L-1 of BAP and 0.5 mg L-1 BAP + 0.5 mg L-1 IAA. The experimental results indicated that pre-treatment with cytokinin BAP (1.0 mg L-1) in germination stage stimulates regeneration process in cotyledons and hypocotyls of melon line 11/9. [ABSTRACT FROM AUTHOR]

Crops drought tolerance is a trait of outmost importance for agriculture especially today when climate change is affecting more the production for food and feed. The scope of this article is to evaluate in vitro drought stress response of Nicotiana tabacum L., “Baladi”. The experiment was set up for four successive stages starting with in vitro seedling development, hypocotyl cultivation, three generations of micropropagation, pre-acclimatization and acclimatization. The effect of abscisic acid (ABA) and/or polyethylene-glycol 6000 (PEG) on tobacco hypocotyl caulogenesis and micropropagation were investigated. Superoxide-dismutases (SODs) and peroxidases (POXs) are more active and different isoforms patterns have been identified compared to the control for cualogenesis. A decrease of internodes length and a higher shoots multiplication rate were observed. However, under PEG treatment plantlets expressed hyperhydration and ceased rooting. Pre-treatments effects study of ABA and/or PEG were finalized in acclimatization phase for 18 tobacco clones. A summary of our results revealed that ABA and/or PEG induce among others a higher oxidative stress compared to the control in the first stage that is not maintained for all clones until acclimatization. Certain clones expressed a lower SOD activity compared to the control during acclimatization but maintaining higher POX activity.

Size control is a fundamental question in biology, showing incremental complexity in plants, whose cells possess a rigid cell wall. The phytohormone auxin is a vital growth regulator with central importance for differential growth control. Our results indicate that auxin-reliant growth programs affect the molecular complexity of xyloglucans, the major type of cell wall hemicellulose in eudicots. Auxin-dependent induction and repression of growth coincide with reduced and enhanced molecular complexity of xyloglucans, respectively. In agreement with a proposed function in growth control, genetic interference with xyloglucan side decorations distinctly modulates auxin-dependent differential growth rates. Our work proposes that auxin-dependent growth programs have a spatially defined effect on xyloglucan’s molecular structure, which in turn affects cell wall mechanics and specifies differential, gravitropic hypocotyl growth.

Vivipary is not a rare finding in papaya fruit. However, this short communication describes the unusual germination of seedling within intra-ovarian ovary. The seedling is attached at two ends of the immature ovary: by radicle at one end, and by elongated tubular structure at the other end. The tubular structure, probably a hypocotyl, mimics an umbilical cord connecting the growing fetus with that of placenta in human embryonic development. [ABSTRACT FROM AUTHOR]

Techniques based on the use of plant protoplasts are a convenient model for better understanding and observing developmental changes in the cells. The establishment of research tools based on protoplasts consists of many steps needed for optimization. Here, we describe the culture of morphogenic callus (MC)- and hypocotyl-derived protoplasts of common (Fagopyrum esculentum Moench) and Tartary (F. tataricum (L.) Gaertn.) buckwheat. Protoplasts embedding in agarose matrix and application of plant hormones, including phytosulfokine (PSK), enable the development of protoplast cultures and plant regeneration., (© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

This study was conducted to compare the effects of NaCl, KCl, MgCl2, MgSO4 and CaCl2 on the seed germination and early seedling growth of MTi2. Five types of salts (NaCl, KCl, MgCl2, MgSO4 and CaCl2) at different concentration (50, 100, 150, and 200 mM) and deionized water as control are used. A10 sterilized seeds were placed in petri dishes containing 5ml of deionized water or each salinity solution and kept in the growth chamber at 25 ± 1°C. The experiment is conducted in a completely randomized design with nine replicates. The number of germinated seeds was recorded daily until day 8. The length hypocotyl and radicle length and the biomass of seedlings were measured at day 8. Germination percentage, seed vigor and salt tolerant were calculated. Data were analyzed using SPSS windows version 22. Data are subjected two way ANOVA at p ≤ 0.05 to determine the significant difference between treatment and followed Tukey at p ≤ 0. 05 for means comparison. Results show the response of MTi2 seed on five different salts is significantly different. It indicates that the viability of MTi2 seed to germinate relatively high in KCl followed by NaCl, CaCl2, MgSO4, and MgCl2. [ABSTRACT FROM AUTHOR]

Light is fundamental for plants in growth and development, with regulating by integration of photoreceptors, hormones, and transcription factors. In this study, the cucumber seedlings were supplemented with different ratios of red and blue light (1R1B, 2R1B, 1R2B and weak daylight as control), with triggering change of phenotype, and gene expression profiles of Cs PHYs and Cs CRYs , Cs GAs / BRs , and CsPIFs in hypocotyls. The cucumber seedling growth was significantly improved by supplemental light quality as compared with CK, and the seedlings in 2R1B were the stoutest, with obviously shortening hypocotyls, and higher dry weight and seedlings index at two-leaf stage. The gene expression of photoreceptor and hormone, including CsPHYA , CsPHYB , CsCRY1 , CsGA20ox1 , CsGA20ox2 , CsGA3ox1 , was significantly up-regulated in hypocotyl under different supplemental light conditions. The cucumber seedlings silenced by pTRV2-PIF4 had an obvious shortened hypocotyl. The expression level of CsCRY1 , CsGA20ox1 and CsGA3ox1 was markedly down-regulation, whereas CsPHYA and CsPHYB expression increased observably and CsGA20ox2 expression was not dramatically difference in pTRV2-PIF4-infected seedlings. Thus, cucumber seedlings hypocotyl elongation was regulated by different supplemental light through crosstalk of photoreceptor, GAs, PIFs, and increasing ratio of red light could promote suppression of hypocotyl elongation. • The seedlings in 2R1B were the stoutest, with shortened hypocotyl, higher dry weight and seedlings index at two-leaf stage. • The cucumber seedlings silenced by pTRV2-PIF4 had an obvious shortened hypocotyl. • In pTRV2-PIF4-infected seedlings, expression of CsCRY1 , CsGA20ox1 and CsGA3ox1 decreased, while CsPHYA and CsPHYB increased. • PIF4 might play a hub role in crosstalk of GA, BR and light signals to regulate photomorphogenesis. [ABSTRACT FROM AUTHOR]

Clusterbean (C. tetragonoloba) is an important, leguminous vegetable and industrial crop with vast genetic diversity but meager genetic, cytological and genomic information. In the present study, an optimized procedure of flow cytometry was used to estimate the genome size of three clusterbean species, represented by C. tetragonoloba (cv. RGC-936) and two wild relatives (C. serreta and C. senegalensis). For accurate estimation of genomic content, singlet G 0 /G 1 populations of multiple tissues such as leaves, hypocotyl, and matured seeds were determined and used along with three different plant species viz. Pisum sativum (as primary), Oryza sativa , and Glycine max (secondary), as external and internal reference standards. Seed tissue of the test sample and G. max provided the best estimate of nuclear DNA content in comparison to other sample tissues and reference standards. The genome size of C. tetragonoloba was detemined at 580.9 ± 0.02 Mbp (1C), while that of C. serreta and C. senegalensis was estimated at 979.6 ± 0.02 Mbp (1C) and 943.4 ± 0.03 Mbp (1C), respectively. Thus, the wild relatives harbor, nearly double the genome content of the cultivated cluster bean. Findings of this study will enrich genomic database of the legume family and can serve as the starting point for clusterbean evolutionary and genomics studies. • During flow cytometry analysis, the seed was found to be the excellent tissue in comparison to leaf and hypocotyl. • Glycine max was found to be a potential reference standard in comparison to other reference standards used. • Significant inter-specific variation while very less inter-tissue variation within a species was observed. • The genome size of C. tetragonoloba , C. senegalenesis and C. serrata was found to be 580.9 Mbp, 943.4 Mbp and 979.6 Mbp respectively. [ABSTRACT FROM AUTHOR]