Botao Song
Huazhong Agricultural University
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Featured researches published by Botao Song.
Molecular Genetics and Genomics | 2011
Xun Liu; Chi Zhang; Yongbin Ou; Yuan Lin; Botao Song; Conghua Xie; Jun Liu; Xiu-Qing Li
Acid invertase is believed to play a regulatory role during plant developmental processes and to respond to environmental stimuli. The expression profiles of the entire acid invertase family are not yet available for potato. By searching existing databases, it was determined that there are at least six acid invertase genes in potato, including four cell-wall invertase genes and two vacuolar invertase genes. They were subjected to comparative expression profiling in various organs of potato plants and in stored tubers to exploit their potential functions. The results revealed that each gene exhibited a unique expression pattern, which differed in transcript abundance or showed organ-specific features, pointing to the possible involvement of individual genes in plant development. The vacuolar invertase gene StvacINV1 had the highest expression level among three genes detected in the potato tubers. Further storage experiments showed that StvacINV1 was strongly induced by low temperatures, which is consistent with glucose accumulation in cold-stored tubers. Suppression of StvacINV1 by the antisense transformation in potato confirmed that lower StvacINV1 transcript abundance in transgenic tubers is related to lower reducing sugar content and lighter chip color in comparison with the wild type. The evidence strongly suggests that StvacINV1 is a gene involved in regulation of cold-induced sweetening of potato tubers. This provides an avenue for studying the mechanism involved in the regulation of the cold-induced sweetening trait and for agronomic enhancement.
Molecular Genetics and Genomics | 2010
Xun Liu; Botao Song; Huiling Zhang; Xiu-Qing Li; Conghua Xie; Jun Liu
Invertase inhibitors (InvInh) interacted with invertases (Inv) and inhibited their activities involved in reducing sugars (RS) accumulation in cold-stored potato tubers. Understanding their potential contribution to RS accumulation is of both theoretical and practical importance because RS accumulation is a costly postharvest problem for both potato producers and processors. In this study, four genes with significant sequence homology to NtInvInhs were identified from potato and their possible contributions to cold-induced sweetening (CIS) of tubers were investigated together with StInv1, an acid invertase gene previously clarified corresponsive to CIS. Transcripts analysis of these StInvInhs and StInv1 among six potato genotypes with distinct CIS sensitivity indicated that StInvInh2 had a negative power regression to RS increase of the cold-stored tubers while a positive linear regression was obtained with StInv1. The relative expression ratio calculated by StInv1/StInvInh2 performed a very significant correlation to RS accumulation, suggesting a possible interaction between StInv1 and StInvInh2 in response to CIS. The bimolecular fluorescence complementation visualized the interaction between StInv1 and StInvInh2A and with StInvInh2B in both onion epidermal cells and tobacco BY-2 cells and demonstrated that these two inhibitors may be the isoforms of StInvInh2 as the counterparts of StInv1. The recombinant StInvInh2B protein inhibited the activities of soluble acid invertase indicating evidently its inhibitory properties. Our results strongly suggest that the interaction between StInv1 and StInvInh2 may play critical roles in controlling the CIS through posttranslational regulation of StInv1 by StInvInh2 in potato tubers and will provide novel tools and resources for improving CIS tolerance of potatoes.
Plant Biotechnology Journal | 2013
Xun Liu; Yuan Lin; Jun Liu; Botao Song; Yongbin Ou; Huiling Zhang; Meng Li; Conghua Xie
Reducing sugar (RS) accumulation in cold-stored potato tubers, known as cold-induced sweetening (CIS), is a crucial factor causing unacceptable colour changes and acrylamide formation of fried products. The activity of vacuolar invertase (StvacINV1) is proved important for the CIS process, and invertase inhibitors are speculated to play roles in the post-translational regulation of StvacINV1 activity. In our previous research, two putative inhibitors (StInvInh2A and StInvInh2B) of StvacINV1 were implied to be involved in potato CIS. Here, we further reported that StInvInh2A and StInvInh2B had similar function that specifically inhibited StvacINV1 activity in potatoes. The genetic transformation of these inhibitor genes in potatoes by overexpression in CIS-sensitive and RNAi-silenced in CIS-resistant genotypes showed that StvacINV1 activity was strongly regulated by alteration of the transcripts of the inhibitors without impacting on the expression of StvacINV1. A negative power relationship was found between the transcripts of the inhibitors and StvacINV1 activity, suggesting 1) a transcriptional determination of the inhibitory capacity of StInvInh2A and StInvInh2B and 2) a significant inhibitory role of these inhibitors in post-translational modulation of StvacINV1. The results also demonstrated that depression of StvacINV1 activity through overexpression of StInvInh2A and StInvInh2B weakened accumulation of RS and acrylamide in cold-stored tubers and consequently improved the chip quality. The present research strongly suggest that both StInvInh2A and StInvInh2B function as inhibitors of StvacINV1 and play similar roles in regulating potato CIS by capping StvacINV1 activity. These inhibitors could be novel genetic resources applicable for improving quality of potato processing products.
Plant Cell Reports | 2007
Qing Zhu; Botao Song; Chi Zhang; Yongbin Ou; Conghua Xie; Jun Liu
The improvement of processing quality of potato products (fries and chips) demands less accumulation of reducing sugars (glucose and fructose) in cold-stored potato (Solanum tuberosum) tubers. Control of gene expression to achieve this requires promoters with specificity to tubers as well as inducible activity under low temperatures. Here we use overlapping extension PCR to construct two chimeric promoters, pCL and pLC, to control gene expression in a tuber-specific and cold-inducible pattern. This combined different combinations of the LTRE (low-temperature responsive element) from Arabidopsis thaliana cor15a promoter and the TSSR (tuber-specific and sucrose-responsive sequence) from potato class I patatin promoter. The cold-inducible and tuber-specific activities of the chimeric promoters were investigated by quantitative analysis of GUS activity in transgenic potato cultivar E3 plants. The results showed that the cis-elements, LTRE and TSSR, played responsive roles individually or in combination. pCL with the TSSR closer to the TATA-box showed substantially higher promoter activity than pLC with the LTRE closer to the TATA-box at either normal (20°C) or low temperature (2°C), suggesting that the promoter activity was closely associated with the position of the two elements. The chimeric promoter pCL with tuber-specific and cold-inducible features may provide valuable tool for controlling the expression of gene constructs designed to lower the formation of reducing sugars in tubers stored at low temperature and to improve the processing quality of potato products.
Journal of Plant Physiology | 2010
Xuemei Ni; Zhendong Tian; Jun Liu; Botao Song; Conghua Xie
Really interesting new gene (RING) finger proteins function as ubiquitin ligase and play key roles in biotic and abiotic stresses. A new RING-H2 finger protein gene, StRFP1, was cloned from Phytophthora infestans-inoculated leaves of potato (Solanum tuberosum) clone 386209.10, which is free of R1-R11 genes. The deduced amino acid sequence was characterized by an N-terminal transmembrane domain, a GLD region and a RING-H2 finger signature. StRFP1 is homologous to the tobacco NtACRE132 protein and belongs to the ATL family. The DNA gel blot analysis and mapping revealed that StRFP1, an intron-free gene, had one to two copies in the potato genome and was located on chromosome 3. RT-PCR assays showed that StRFP1 was constitutively expressed in potato plants and significantly induced in detached potato leaves by P. infestans and plant defense-related signal molecules, abscisic acid, salicylic acid and methyl jasmonate. Transient expression studies revealed that StRFP1 fused with GFP localized to the plasma membrane or out of that in onion epidermal cells. The function of StRFP1 in potato resistance against late blight was further investigated by constructing overexpression and RNA interference (RNAi) vectors, which were introduced into potato cv. E-potato 3, respectively. By challenging the detached leaves with mixture races of P. infestans, all of the StRFP1-overexpressing plants displayed slower disease development than non-transformed controls in terms of the lesion growth rate (LGR). In contrast, StRFP1-silencing plants through RNAi were more susceptible to pathogen infection. The present results demonstrate that StRFP1 contributes to broad-spectrum resistance against P. infestans in potato.
Theoretical and Applied Genetics | 2012
Jingcai Li; Hannele Lindqvist-Kreuze; Zhendong Tian; Jun Liu; Botao Song; Juan Landeo; Leticia Portal; Manuel Gastelo; Julio Frisancho; Laura Sanchez; Dennis Meijer; Conghua Xie; Merideth Bonierbale
A large number of quantitative trait loci (QTL) for resistance to late blight of potato have been reported with a “conventional” method in which each phenotypic trait reflects the cumulative genetic effects for the duration of the disease process. However, as genes controlling response to disease may have unique contributions with specific temporal features, it is important to consider the phenotype as dynamic. Here, using the net genetic effects evidenced at consecutive time points during disease development, we report the first conditional mapping of QTL underlying late blight resistance in potato under five environments in Peru. Six conditional QTL were mapped, one each on chromosome 2, 7 and 12 and three on chromosome 9. These QTL represent distinct contributions to the phenotypic variation at different stages of disease development. By comparison, when conventional mapping was conducted, only one QTL was detected on chromosome 9. This QTL was the same as one of the conditional QTL. The results imply that conditional QTL reflect genes that function at particular stages during the host–pathogen interaction. The dynamics revealed by conditional QTL mapping could contribute to the understanding of the molecular mechanism of late blight resistance and these QTL could be used to target genes for marker development or manipulation to improve resistance.
Molecular Genetics and Genomics | 2012
Xia Chen; Botao Song; Jun Liu; Jianwen Yang; Tianjiu He; Yuan Lin; Huiling Zhang; Conghua Xie
Cold-induced sweetening (CIS) is a crucial factor influencing the processing quality of potato tubers. To better understand the molecular events of potato CIS and different CIS-sensitivity among various potato species, a suppression subtractive hybridization library and cDNA microarray gene filters were developed. A total of 188 genes were found to be differentially expressed (DE) in Solanum berthaultii (ber) upon cold stimulation. These functional genes were mostly related to cell rescue, defense and virulence, metabolism, energy and protein fate, included in various processes of plant defense against abiotic stresses. Four expression patterns of these DE genes were profiled by qRT-PCR using the cold-stored tubers of both CIS-resistant (ber) and CIS-sensitive (E-potato 3, a variety of S. tuberosum) potatoes. The expression pattern and abundance of many DE genes encoding proteins involved in metabolism were different in these two potato tubers, especially genes associated with amylolysis, sucrose decomposition and glycolysis pathways, indicating distinct regulatory mechanisms between ber and E3 in response to cold stress, which may be crucial for potato CIS. Further investigation of these cold-regulated genes will deepen our understanding of the regulatory mechanisms of potato CIS and direct approaches for the genetic improvement of potato processing quality.
Plant Physiology | 2015
Yuan Lin; Tengfei Liu; Jun Liu; Xun Liu; Yongbin Ou; Huiling Zhang; Meng Li; Uwe Sonnewald; Botao Song; Conghua Xie
Invertase activity is affected by a protein complex to modulate accumulation of reducing sugars in cold-stored potato tubers. Slowing down cold-induced sweetening (CIS) of potato (Solanum tuberosum) tubers is of economic importance for the potato industry to ensure high-quality products. The conversion of sucrose to reducing sugars by the acid invertase StvacINV1 is thought to be critical for CIS. Identification of the specific StvacINV1 inhibitor StInvInh2B and the α- and β-subunits of the interacting protein SUCROSE NONFERMENTING1-RELATED PROTEIN KINASE from the wild potato species Solanum berthaultii (SbSnRK1) has led to speculation that invertase activity may be regulated via a posttranslational mechanism that remains to be elucidated. Using bimolecular fluorescence complementation assays, this study confirmed the protein complex by pairwise interactions. In vitro kinase assays and protein phosphorylation analysis revealed that phosphorylation of SbSnRK1α is causal for StvacINV1 activity and that its active form blocks the inhibition of StInvInh2B by SbSnRK1β, whereas its inactive form restores the function of SbSnRK1β that prevents StInvInh2B from repressing StvacINV1. Overexpression of SbSnRK1α in CIS-sensitive potato confirmed that SbSnRK1α has significant effects on acid invertase-associated sucrose degradation. A higher level of SbSnRK1α expression was accompanied by elevated SbSnRK1α phosphorylation, reduced acid invertase activity, a higher sucrose-hexose ratio, and improved chip color. Our results lend new insights into a subtle regulatory mode of invertase activity and provide a novel approach for potato CIS improvement.
Plant Cell Reports | 2010
Xianpu Guo; Conghua Xie; Xingkui Cai; Botao Song; Li He; Jun Liu
Potato somatic hybrids obtained by protoplast fusion between Solanum tuberosum (4x) and Solanum chacoense (2x) were investigated for genome stability and meiotic behavior associated with the pollen viability in order to elucidate the mechanism influencing the fertility of the somatic hybrids. The ploidy level detections conducted in 2004 and 2007 demonstrated that 68 out of 108 somatic hybrids had their ploidy level changed to be uniform and euploidy after successive in vitro subcultures, which mainly occurred in octaploids, aneuploids, and mixoploids, while 74% hexaploids were still stable in their genome dosage in 2007. Different types of abnormal meiotic behavior were observed during the development of pollen mother cells (PMCs) including the formation of univalents, multivalents, laggard chromosomes, and chromosomal bridges, as well as triads and polyads. A higher proportion of abnormal meiosis seemed to be accompanied with a genome dosage higher than the hexaploids expected in this study. A significant positive correlation between defective PMCs and the number of small pollen grains and negative correlation between number of small pollen grains and pollen viability strongly suggested that abnormal meiosis could be a causal factor influencing the fertility of the somatic hybrids. The hexaploids with stable genome dosage and a certain level of fertility will have great potential in a potato breeding program.
FEBS Letters | 2013
Huiling Zhang; Xun Liu; Jun Liu; Yongbin Ou; Yuan Lin; Meng Li; Botao Song; Conghua Xie
The modulation of the activity of enzymes associated with carbohydrate metabolism is important for potato cold‐induced sweetening (CIS). A novel RING finger gene SbRFP1 was cloned and its expression was found to be cold‐inducible in potato tubers of the CIS‐resistant genotypes. Transformation of SbRFP1 in potatoes confirmed its role in inhibiting β‐amylase and invertase activity, which consequently slowed down starch and sucrose degradation and the accumulation of reducing sugars in cold stored tubers. These findings strongly suggest that SbRFP1 may function as a negative regulator of BAM1 and StvacINV1 to decelerate the accumulation of reducing sugars in the process of potato CIS.