Browsing by Author "Mei, Chuansheng"
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- Burkholderia phytofirmans strain PsJN effects on drought resistance, physiological responses and growth of switchgrassWang, Bingxue (Virginia Tech, 2015-02-09)To decrease dependency of fossil fuels and avoid direct competition with food crops, massive research efforts are investigating next-generation cellulose biofuel crops such as switchgrass (Panicum virgatum). A low-input, sustainable switchgrass production could be achieved by reducing traditional management practices though applying plant growth promoting rhizobacteria (PGPR), of which our understanding is still rather limited. To elucidate physiological mechanisms behind PGPR's beneficial effects, we inoculated switchgrass seedlings with Burkholderia phytofirmans strain PsJN. Two experiments were conducted to determine the initial and long-term responses of switchgrass to PsJN inoculation by tracking growth and leaf physiology. In a third experiments, we tested the effects of PsJN on growth and leaf-level physiology of switchgrass under a moderate pre-drought conditioning and a successive severe drought stress. PsJN inoculation increased biomass and promoted elongation of shoots within 17 days following inoculation. The enhanced root growth in PsJN inoculated plants lagged behind the shoot response, resulting in greater allocation to aboveground growth (p=0.0041). Lower specific root length (p=0.0158) and higher specific leaf weight (p=0.0029) were also observed in PsJN inoculated seedlings, indicating advanced development. Photosynthetic rates (Ps) were higher in PsJN inoculated seedlings after 17 days (54%, p=0.0016), which were related to higher stomatal conductance, greater water use efficiency, and lower non-stomatal limitation of Ps. These rapid changes in leaf physiology are at least partially responsible for switchgrass growth enhancement from PsJN treatment. The early growth enhancement in PsJN inoculated switchgrass linearly decreased with plant age. PsJN inoculation increased Ps of upper canopy leaves by 13.6% but reduced Ps of lower canopy leaves by 8.2%. Accelerated leaf senescence and early flowering were observed in PsJN-inoculated switchgrass, which might contribute to slightly lower aboveground biomass at final harvesting. Drought preconditioning increased Ps of PsJN-inoculated switchgrass during a later severe drought; whereas, control switchgrass only benefited from drought preconditioning when leaf water potential dropped below -1 MPa. This study verified early growth enhancement and accelerated development of switchgrass due to PsJN inoculation. Rapid improvement in leaf physiology is related to enhanced productivity. PsJN inoculation also improve drought tolerance of switchgrass.
- Characterization of activation tagged potato (Solanum tuberosum L.) mutantsAulakh, Sukhwinder Singh (Virginia Tech, 2012-09-14)Generation and characterization of activation tagged potato mutants could aid in functional genomic studies. Morphological and molecular studies were conducted to compare potato cv. Bintje, its two mutants, underperformer (up), and nikku generated using the activation tagging vector pSKI074, and nikku revertant plants. Mutant up exhibited a dwarf phenotype (plant height 42 cm vs. 73 cm in cv. Bintje), abundant axillary shoot growth (3.1 shoots/plant compared to 0.7 shoots/plant in cv. Bintje; in vitro plants), greater tuber yield, altered tuber traits and early senescence compared to wild-type Bintje under in vitro conditions. Under in vivo conditions, the dwarf and early senescence phenotypes of the mutant were consistent, but the tuber yield of up was less (250 g/plant compared to 610 g/plant in wild-type Bintje) and had fewer axillary shoots compared to wild-type (1.9 shoots/plant in up vs. 4.7 shoots/plant in Bintje). Mutant nikku plants exhibited an extremely dwarf phenotype (plant height 2 cm in nikku vs. 6 cm in Bintje), had small hyponastic leaves, were rootless, and infrequently produced small tubers when compared to cv. Bintje. The overall nikku phenotype was suggestive of a constitutive stress response, which was further supported by the higher expression levels of several stress-responsive genes in nikku. The nikku revertant plants exhibited near normal stem elongation, larger leaves and consistent rooting, and it was a case of partial reversion. Southern blot analyses indicated the presence of single T-DNA insertions on chromosome 10 in the up and on chromosome 12 in the nikku mutant. The reversion in the nikku plants was not associated with the loss of enhancer copies from the original nikku mutant. Reverse transcriptase PCR analyses indicated transcriptional activation/repression of several genes in the up and nikku mutants, suggesting pleiotropic effects. In revertant, the expression levels of several genes which were differentially regulated in the nikku mutant were similar to Bintje. The gene immediately flanking the right border of the T-DNA insertion, which encoded a novel BTB/POZ (Broad complex, Tramtrac, Bric a brac; also known as Pox virus and Zinc finger) domain-containing protein, was highly up-regulated in the up mutant. This protein domain plays an important role in several important developmental, transcriptional and regulatory pathways. The mRNA-seq analyses resulted in 1,632 genes that were differentially expressed between mutant up and Bintje and the total number of up-regulated genes (661) were less than the number of genes down-regulated (971 genes) in the up mutant. Further analyses indicated that a variety of biological processes including decreased cell division, cell cycle activity, and abiotic stress responses were modified in the up mutant. In the nikku mutant, two potato genes, encoding an Acyl-CoA N-acyltransferases (NAT) superfamily protein, and a predicted major facilitator superfamily protein (MFS) were identified and overexpression lines Bintje/35S::NAT1 and Bintje/35S::PMT1 were created for recapitulation of the nikku mutant phenotype. Methylated DNA-PCR between the nikku and the revertant indicated a change in methylation status of the 35S enhancers, suggesting that the nikku revertant phenotype may be associated with some epigenetic modification.
- Genetic Improvement of Switchgrass Cell Wall Content, Leaf Angle and Flowering TimeXu, Bin (Virginia Tech, 2011-06-10)Switchgrass (Panicum virgatum L.) is a candidate bioenergy crop. Somatic embryogenic (SE) calli are used for genetic transformation in switchgrass. A superior switchgrass line, HR8, was developed using recurrent tissue culture selection from cv. Alamo. HR8 SE calli were genetically transformable using Agrobacterium at an efficiency of ~12%. We used HR8 somatic embryogenic calli for genetic improvement of switchgrass. The lignin content of feedstock has been proposed as one key trait impacting biofuel production. 4-Coumarate: Coenzyme A ligase (4CL) is one of the key enzymes involved in the monolignol biosynthetic pathway. Two homologous 4CL genes, Pv4CL1 and Pv4CL2, were identified in switchgrass. Gene expression patterns and enzymatic activity assays suggested that Pv4CL1 is involved in monolignol biosynthesis. Stable transgenic plants were obtained with Pv4CL1 down-regulated. RNA interference of Pv4CL1 reduced extractable 4CL activity by 80%, leading to a reduction in lignin content with decreased guaiacyl unit composition. The transgenic plants had uncompromised biomass yield. After dilute acid pretreatment, the low lignin transgenic biomass had significantly increased cellulose hydrolysis (saccharification) efficiency for biofuel production. Erect leaf is a desirable trait to adjust the overall plant architecture to perceive more solar energy and thereby to increase the plant biomass production in a field population. We overexpressed an Arabidopsis NAC transcriptional factor gene, LONG VEGETATIVE PHASE ONE (AtLOV1), in switchgrass. Surprisingly, AtLOV1 induced smaller leaf angle by changing morphologies of epidermal cells in the leaf collar region, affecting lignin content and monolignol composition, and also causing delayed flowering time in switchgrass. Global gene-expression analysis of AtLOV1 transgenic plants demonstrated an array of genes has altered expressions. Potential downstream genes involved in the pleiotropic phenotypic traits of the transgenic plants are discussed.
- Growth promotion and colonization of switchgrass (Panicum virgatum) cv. Alamo by bacterial endophyte Burkholderia phytofirmans strain PsJNKim, Seonhwa; Lowman, James Scott; Hou, Guichuan; Nowak, Jerzy; Flinn, Barry; Mei, Chuansheng (2012-05-30)Background Switchgrass is one of the most promising bioenergy crop candidates for the US. It gives relatively high biomass yield and can grow on marginal lands. However, its yields vary from year to year and from location to location. Thus it is imperative to develop a low input and sustainable switchgrass feedstock production system. One of the most feasible ways to increase biomass yields is to harness benefits of microbial endophytes. Results We demonstrate that one of the most studied plant growth promoting bacterial endophytes, Burkholderia phytofirmans strain PsJN, is able to colonize and significantly promote growth of switchgrass cv. Alamo under in vitro, growth chamber, and greenhouse conditions. In several in vitro experiments, the average fresh weight of PsJN-inoculated plants was approximately 50% higher than non-inoculated plants. When one-month-old seedlings were grown in a growth chamber for 30 days, the PsJN-inoculated Alamo plants had significantly higher shoot and root biomass compared to controls. Biomass yield (dry weight) averaged from five experiments was 54.1% higher in the inoculated treatment compared to non-inoculated control. Similar results were obtained in greenhouse experiments with transplants grown in 4-gallon pots for two months. The inoculated plants exhibited more early tillers and persistent growth vigor with 48.6% higher biomass than controls. We also found that PsJN could significantly promote growth of switchgrass cv. Alamo under sub-optimal conditions. However, PsJN-mediated growth promotion in switchgrass is genotype specific. Conclusions Our results show B. phytofirmans strain PsJN significantly promotes growth of switchgrass cv. Alamo under different conditions, especially in the early growth stages leading to enhanced production of tillers. This phenomenon may benefit switchgrass establishment in the first year. Moreover, PsJN significantly stimulated growth of switchgrass cv. Alamo under sub-optimal conditions, indicating that the use of the beneficial bacterial endophytes may boost switchgrass growth on marginal lands and significantly contribute to the development of a low input and sustainable feedstock production system.
- A Potential Application of Endophytic Bacteria in Strawberry ProductionMei, Chuansheng; Amaradasa, B. Sajeewa; Chretien, Robert L.; Liu, Danyang; Snead, George; Samtani, Jayesh B.; Lowman, Scott (MDPI, 2021-11-18)Endophytic bacteria could be used as a tool to promote plant growth, enhance abiotic stress tolerance, and inhibit plant pathogen growth. More than 100 bacteria, previously tested to have biocontrol activity against fungal pathogens from our lab, were chosen for pathogen inhibition in vitro with antibiosis assays against strawberry specific pathogens. Three potential endophytic bacteria were tested in the greenhouse for disease reduction and growth performance. Finally, field trials were conducted for fruit yield production. Our results showed that three endophytic bacteria significantly inhibited the growth of the strawberry pathogen Colletotrichum gloeosporioides from antibiosis assays and were identified as Bacillus velezensis strains IALR308, IALR585, and IALR619. Inoculation of these bacteria significantly reduced strawberry disease in greenhouse conditions with pathogen infection. Field trials showed that IALR619 has potential to influence marketable fruit yield when strawberry plants were inoculated twice over the growing season. All three bacteria had the ability to produce auxin and to solubilize phosphate. The antibiotics surfactin and iturin were also detected in IALR585 and IALR619. In conclusion, Bacillus velezensis IALR619 has potential inhibition of strawberry pathogen growth in the greenhouse and possible ability to increase marketable fruit yield in the field.
- A Potential Application of Pseudomonas psychrotolerans IALR632 for Lettuce Growth Promotion in HydroponicsMei, Chuansheng; Zhou, Dongfang; Chretien, Robert L.; Turner, Amy; Hou, Guichuan; Evans, Michael R.; Lowman, Scott (MDPI, 2023-02-02)Controlled environment agriculture hydroponic systems grow plants year-round without restriction from outside environmental conditions. In order to further improve crop yield, plant growth-promoting bacteria were tested on hydroponically grown lettuce (Lactuca sativa) plants. From our bacterial endophyte library, we found one bacterium, Pseudomonas psychrotolerans IALR632, that is promising in promoting lettuce growth in multiple hydroponic systems. When Green Oakleaf lettuce seeds were inoculated with IALR632 during germination, IALR632 significantly increased lateral root development by 164%. When germinated seedlings were inoculated with IALR632 and then transplanted to different hydroponic systems, shoot and root fresh weights of Green Oakleaf increased by 55.3% and 17.2% in a nutrient film technique (NFT) system in the greenhouse, 13.5% and 13.8% in an indoor vertical NFT system, and 15.3% and 13.6% in a deep water cultivation (DWC) system, respectively. IALR632 also significantly increased shoot fresh weights of Rex by 33.9%, Red Oakleaf by 21.0%, Red Sweet Crisp by 15.2%, and Nancy by 29.9%, as well as Red Rosie by 8.6% (no significant difference). Inoculation of IALR632-GFP and subsequent analysis by confocal microscopy demonstrated the endophytic nature and translocation from roots to shoots. The results indicate that P. psychrotolerans IALR632 has a potential application in hydroponically grown lettuce plants.
- Utilizing Beneficial Bacterial Endophytes to Promote Switchgrass Growth in Low- input Agricultural Production SystemsLowman, James Scott (Virginia Tech, 2014-02-20)The US Department of Energy has focused research efforts on developing switchgrass into a bioenergy feedstock, helping to offset the use of non-renewable fossil fuels and make the US more energy independent. Bacterial endophytes, which reside inside plant tissues, are proven to increase yield and stress resistance in a number of plants. The primary objective of this dissertation was to explore the use of endophytes to improve biomass yields of switchgrass on lands not suitable for food crops and better understand the underlying mechanisms of the plant-endophyte interaction. Integration of this research into K-12 STEM education to increase interest in plant sciences and create the next generation of scientists with the motivation to help solve the challenges facing society in the twenty first century was the objective of the outreach component of this project. Chapter one demonstrates the ability of Burkholderia phytofirmans strain PsJN to colonize switchgrass and promote plant growth under in vitro (approximately 50% higher), and growth chamber and greenhouse (48.6% higher biomass yields) conditions. The objectives of Chapter two were to determine stand establishment in the field with different nutrient levels. PsJN bacterization positively benefited growth and development of switchgrass seedlings in the field with both low and high nutrient content. Highly significant (p<0.001) stimulation of root and shoot growth, lateral root formation and number of tillers was recorded on soil with low fertility. PsJN bacterization also enhanced biomass accumulation during the two seasons of growth on both poor (p<0.001) and rich (p<0.05) soil, indicating the potential for the use of PsJN in a low-input switchgrass feedstock production system. Chapter three outlines differences in gene expression patterns upon bacterization, between the responsive cv. Alamo, and a non-responsive cv. Cave-in-Rock. Using EST microarrays and quantitative PCR up- and down-regulated genes were identified in both cultivars. One of the key genes identified was a member of the tau class, glutathione S-transferase (GST). GST enzymes are known to be involved in plants responses to stress. Using overexpression and knockout/knockdown techniques we demonstrated that GST is likely involved in the bacterization induced early plant growth promotion in switchgrass. Chapter four describes the potential for the utilization of beneficial bacterial endophytes capable of fixing atmospheric nitrogen in a free-living state in the development of low-input switchgrass feedstock production systems. Sphingomonas sp. strain NSL isolated from switchgrass tissue was able to grow on nitrogen free medium and stimulated growth of switchgrass cv. Alamo under nitrogen deficient conditions. The ability to fix atmospheric nitrogen was also moved to Burkholderia phytofirmans strain PsJN via horizontal gene transfer from the legume nodulating Burkholderia phymatum. The transformed PsJN was able to fix nitrogen and promote plant growth under nitrogen limited conditions. At every step of the research described in this dissertation efforts were made to include its elements into K-12 education. Chapter five describes four case studies aiming at the enhancement of youth interest in plant sciences in the socieoeconomically depressed areas of Southside Virginia.