Starch is the predominant compound in bulb scales,and previous studies have shown that bulblet development is closely associated with starch enrichment.However,how starch synthesis affects bulbification at the molecul...Starch is the predominant compound in bulb scales,and previous studies have shown that bulblet development is closely associated with starch enrichment.However,how starch synthesis affects bulbification at the molecular level is unclear.In this study,we demonstrate that Lilium brownii var.giganteum,a wild lily with a giant bulb in nature,and L.brownii,the native species,have different starch levels and characteristics according to cytological and ultra-structural observations.We cloned the complete sequence of three key gene-encoding enzymes(LbgAGPS,LbgGBSS,and LbgSSⅢ)during starch synthesis by rapid amplification of 5’and 3’complementary DNA(cDNA)ends(RACE)technology.Bioinformatics analysis revealed that the proteins deduced by these genes contain the canonical conserved domains.Constructed phylogenetic trees confirmed the evolutionary relationships with proteins from other species,including monocotyledons and dicotyledons.The transcript levels of various tissues and time course samples obtained during bulblet development uncovered relatively high expression levels in bulblets and gradual increase expression accompanying bulblet growth.Moreover,a set of single nucleotide polymorphisms(SNPs)was discovered in the AGPS genes of four lily genotypes,and a purifying selection fashion was predicted according to the non-synonymous/synonymous(Ka/Ks)values.Taken together,our results suggested that key starch-synthesizing genes might play important roles in bulblet development and lead to distinctive phenotypes in bulblet size.展开更多
Bulblet development is a problem in global lily bulb production and carbohydrate metabolism is a crucial factor. Micropropagation acts as an efficient substitute for faster propagation and can provide a controllable c...Bulblet development is a problem in global lily bulb production and carbohydrate metabolism is a crucial factor. Micropropagation acts as an efficient substitute for faster propagation and can provide a controllable condition to explore bulb growth. The present study was conducted to investigate the effects of humic acid （HA） on bulblet swelling and the carbohydrate metabolic pathway in Li/ium Oriental Hybrids ＇Sorbonne＇ under in vitro conditions. HA greatly promoted bulblet growth at 0.2, 2.0, and 20.0 mg/L, and pronounced increases in bulblet sucrose, total soluble sugar, and starch content were observed for higher HA concentrations （_〉2.0 mg/L） within 45 d after transplanting （DAT）. The activities of three major starch synthetic enzymes （including adenosine 5＇-diphosphate glucose pyro- phosphorylase, granule-bound starch synthase, and soluble starch synthase） were enhanced dramatically after HA application especially low concentration HA （LHA）, indicating a quick response of starch metabolism. However, higher doses of HA also caused excessive aboveground biomass accumulation and inhibited root growth. Accordingly, an earlier carbon starvation emerged by observing evident starch degradation. Relative bulblet weight gradually decreased with increased HA doses and thereby broke the balance between the source and sink. A low HA concentration at 0.2 mg/L performed best in both root and bulblet growth. The number of roots and root length peaked at 14.5 and 5.75 cm respectively. The fresh bulblet weight and diameter reached 468 mg （2.9 times that under the control treatment） and 11.68 mm, respectively. Further, sucrose/starch utilization and conversion were accelerated and carbon famine was delayed as a result with an average relative bulblet weight of 80.09%. To our knowledge, this is the first HA application and mechanism research into starch metabolism in both in vitro and in vivo condition in bulbous crops.展开更多
Lily(Lilium spp.) is an important ornamental flower, which is mainly propagated by bulbs. Cell wall invertases(CWINs), which catalyze the irreversibly conversion of sucrose into glucose and fructose in the extracellul...Lily(Lilium spp.) is an important ornamental flower, which is mainly propagated by bulbs. Cell wall invertases(CWINs), which catalyze the irreversibly conversion of sucrose into glucose and fructose in the extracellular space, are key enzymes participating in sucrose allocation in higher plants. Previous studies have shown that CWINs play an essential role in bulblet initiation process in bulbous crops, but the underlying molecular mechanism remains unclear. Here, a CWIN gene of Lilium brownii var. giganteum(Lbg) was identified and amplified from genomic DNA. Quantitative RT-PCR assays revealed that the expression level of LbgCWIN1 was highly upregulated exactly when the endogenous starch degraded in non-sucrose medium during in vitro bulblet initiation in Lbg. Phylogenetic relationship, motif, and domain analysis of LbgCWIN1 protein and CWINs in other plant species showed that all sequences of these CWIN proteins were highly conserved. The promoter sequence of LbgCWIN1 possessed a number of alpha-amylase-, phytohormone-, light-and stress-responsive cis-elements. Meanwhile, β-glucuronidase(GUS) assay showed that the 459 bp upstream fragment from the translational start site displayed maximal promoter activity. These results revealed that LbgCWIN1 might function in the process of in vitro bulblet initiation and be in the response to degradation of endogenous starch.展开更多
基金the National Natural Science Foundation of China(No.31772337)the Lishui Research Fund(No.2020zdhz03)+1 种基金the China Postdoctoral Science Foundation(No.2017M612005)the National Key Research and Development Program of China(No.2018YFD1000401)。
文摘Starch is the predominant compound in bulb scales,and previous studies have shown that bulblet development is closely associated with starch enrichment.However,how starch synthesis affects bulbification at the molecular level is unclear.In this study,we demonstrate that Lilium brownii var.giganteum,a wild lily with a giant bulb in nature,and L.brownii,the native species,have different starch levels and characteristics according to cytological and ultra-structural observations.We cloned the complete sequence of three key gene-encoding enzymes(LbgAGPS,LbgGBSS,and LbgSSⅢ)during starch synthesis by rapid amplification of 5’and 3’complementary DNA(cDNA)ends(RACE)technology.Bioinformatics analysis revealed that the proteins deduced by these genes contain the canonical conserved domains.Constructed phylogenetic trees confirmed the evolutionary relationships with proteins from other species,including monocotyledons and dicotyledons.The transcript levels of various tissues and time course samples obtained during bulblet development uncovered relatively high expression levels in bulblets and gradual increase expression accompanying bulblet growth.Moreover,a set of single nucleotide polymorphisms(SNPs)was discovered in the AGPS genes of four lily genotypes,and a purifying selection fashion was predicted according to the non-synonymous/synonymous(Ka/Ks)values.Taken together,our results suggested that key starch-synthesizing genes might play important roles in bulblet development and lead to distinctive phenotypes in bulblet size.
基金Project supported by the National High-Tech R&D Program(863) of China(No.2011AA100208)the Zhejiang Provincial Natural Science Foundation of China(No.LY12C15003)
文摘Bulblet development is a problem in global lily bulb production and carbohydrate metabolism is a crucial factor. Micropropagation acts as an efficient substitute for faster propagation and can provide a controllable condition to explore bulb growth. The present study was conducted to investigate the effects of humic acid （HA） on bulblet swelling and the carbohydrate metabolic pathway in Li/ium Oriental Hybrids ＇Sorbonne＇ under in vitro conditions. HA greatly promoted bulblet growth at 0.2, 2.0, and 20.0 mg/L, and pronounced increases in bulblet sucrose, total soluble sugar, and starch content were observed for higher HA concentrations （_〉2.0 mg/L） within 45 d after transplanting （DAT）. The activities of three major starch synthetic enzymes （including adenosine 5＇-diphosphate glucose pyro- phosphorylase, granule-bound starch synthase, and soluble starch synthase） were enhanced dramatically after HA application especially low concentration HA （LHA）, indicating a quick response of starch metabolism. However, higher doses of HA also caused excessive aboveground biomass accumulation and inhibited root growth. Accordingly, an earlier carbon starvation emerged by observing evident starch degradation. Relative bulblet weight gradually decreased with increased HA doses and thereby broke the balance between the source and sink. A low HA concentration at 0.2 mg/L performed best in both root and bulblet growth. The number of roots and root length peaked at 14.5 and 5.75 cm respectively. The fresh bulblet weight and diameter reached 468 mg （2.9 times that under the control treatment） and 11.68 mm, respectively. Further, sucrose/starch utilization and conversion were accelerated and carbon famine was delayed as a result with an average relative bulblet weight of 80.09%. To our knowledge, this is the first HA application and mechanism research into starch metabolism in both in vitro and in vivo condition in bulbous crops.
基金financially supported by the National Natural Science Foundation of China (Grant Nos.32101571,32002071)the Zhejiang Science and Technology Major Program on Agricultural New Variety Breeding (Grant No.2021C02071-6)。
文摘Lily(Lilium spp.) is an important ornamental flower, which is mainly propagated by bulbs. Cell wall invertases(CWINs), which catalyze the irreversibly conversion of sucrose into glucose and fructose in the extracellular space, are key enzymes participating in sucrose allocation in higher plants. Previous studies have shown that CWINs play an essential role in bulblet initiation process in bulbous crops, but the underlying molecular mechanism remains unclear. Here, a CWIN gene of Lilium brownii var. giganteum(Lbg) was identified and amplified from genomic DNA. Quantitative RT-PCR assays revealed that the expression level of LbgCWIN1 was highly upregulated exactly when the endogenous starch degraded in non-sucrose medium during in vitro bulblet initiation in Lbg. Phylogenetic relationship, motif, and domain analysis of LbgCWIN1 protein and CWINs in other plant species showed that all sequences of these CWIN proteins were highly conserved. The promoter sequence of LbgCWIN1 possessed a number of alpha-amylase-, phytohormone-, light-and stress-responsive cis-elements. Meanwhile, β-glucuronidase(GUS) assay showed that the 459 bp upstream fragment from the translational start site displayed maximal promoter activity. These results revealed that LbgCWIN1 might function in the process of in vitro bulblet initiation and be in the response to degradation of endogenous starch.
