scholarly journals CRISPR/Cas9-mediated targeted mutagenesis of GmLCL genes alters plant height and internode length in soybean

2019 ◽  
Author(s):  
Fanjiang Kong ◽  
Qun Cheng ◽  
Lidong Dong ◽  
Tong Su ◽  
Zhuoran Gan ◽  
...  

Abstract Background: Soybean ( Glycine max ) is an important economically crops for plant oil and protein in the world. The plant height as a key trait has significant effects on yield of soybean , however, the research on molecular mechanism for soybean plant height is still unclear. Recently, CRISPR (clustered regularly interspaced short palindromic repeat)/Cas9 (CRISPR-associated) system as a new technology for gene editing, has been rapidly utilized to edit the genomes of crop plants. Results: Here, we designed four gRNAs to mutate four LATE ELONGATED HYPOCOTYL ( LHY ) / CIRCADIAN CLOCK ASSOCIATED1 ( CCA1 )- LIKE ( LCL ) genes in soybean. In order to test whether the gRNAs could perform properly in transgenic soybean plants, we first tested the CRISPR construct in transgenic soybean hairy roots using Agrobacterium rhizogenesis strain K599. Once confirmed, we performed stable soybean transformation and obtained nineteen independent transgenic soybean plants. Subsequently, we obtained one T 1 transgene-free homozygous quadruple mutant of GmLCL by self-crossed. The phenotype of T 2 -generation transgene-free quadruple mutant plants were observed and the results showed that quadruple mutant of GmLCL displayed reduced plant height and shortened internodes. In addition, the relative expression levels of gibberellic acid (GA) metabolic pathway genes in the quadruple mutant of GmLCL were significantly decreased than wild type (WT). It suggests that GmLCLs encoding MYB transcription factor affect plant height through mediating the GA pathway in soybean. We also develop some genetic markers to identify mutant for assisting breeding studies. Conclusions: Our results indicate that CRISPR/Cas9-mediated targeted mutagenesis of four GmLCL genes reduce soybean plant height and shorten internodes. These findings suggest that manipulation of four GmLCL genes may improve yield by altered plant height and internode length in soybean.

2019 ◽  
Author(s):  
Fanjiang Kong ◽  
Qun Cheng ◽  
Lidong Dong ◽  
Tong Su ◽  
Zhuoran Gan ◽  
...  

Abstract Background: Soybean (Glycine max) is an economically important oil and protein crop. Plant height is a key trait that significantly impacts the yield of soybean; however, research on the molecular mechanisms associated with soybean plant height is lacking. The CRISPR (clustered regularly interspaced short palindromic repeat)/Cas9 (CRISPR-associated) system is a recently developed technology for gene editing that has been utilized to edit the genomes of crop plants. Results: Here, we designed four gRNAs to mutate four LATE ELONGATED HYPOCOTYL ( LHY ) / CIRCADIAN CLOCK ASSOCIATED1 ( CCA1 )- LIKE ( LCL ) genes in soybean. In order to test whether the gRNAs could perform properly in transgenic soybean plants, we first tested the CRISPR construct in transgenic soybean hairy roots using Agrobacterium rhizogenesis strain K599. Once confirmed, we performed stable soybean transformation and obtained nineteen independent transgenic soybean plants. Subsequently, we obtained one T 1 transgene-free homozygous quadruple mutant of GmLCL by self-crossed. The phenotype of T 2 -generation transgene-free quadruple mutant plants were observed and the results showed that quadruple mutant of GmLCL displayed reduced plant height and shortened internodes. In addition, the relative expression levels of gibberellic acid (GA) metabolic pathway genes in the quadruple mutant of GmLCL were significantly decreased than wild type (WT). It suggests that GmLCLs encoding MYB transcription factor affect plant height through mediating the GA pathway in soybean. We also develop some genetic markers to identify mutant for assisting breeding studies. Conclusions: Our results indicate that CRISPR/Cas9-mediated targeted mutagenesis of four GmLCL genes reduce soybean plant height and shorten internodes. These findings suggest that manipulation of four GmLCL genes may improve yield by altered plant height and internode length in soybean.


2019 ◽  
Author(s):  
Qun Cheng ◽  
Lidong Dong ◽  
Tong Su ◽  
Tingyu Li ◽  
Zhuoran Gan ◽  
...  

Abstract Background: Soybean (Glycine max) is an economically important oil and protein crop. Plant height is a key trait that significantly impacts the yield of soybean; however, research on the molecular mechanisms associated with soybean plant height is lacking. The CRISPR (clustered regularly interspaced short palindromic repeat)/Cas9 (CRISPR-associated system 9) system is a recently developed technology for gene editing that has been utilized to edit the genomes of crop plants. Results: Here, we designed four gRNAs to mutate four LATE ELONGATED HYPOCOTYL (LHY) genes in soybean. In order to test whether the gRNAs could perform properly in transgenic soybean plants, we first tested the CRISPR construct in transgenic soybean hairy roots using Agrobacterium rhizogenes strain K599. Once confirmed, we performed stable soybean transformation and obtained 19 independent transgenic soybean plants. Subsequently, we obtained one T1 transgene-free homozygous quadruple mutant of GmLHY by self-crossing. The phenotypes of the T2-generation transgene-free quadruple mutant plants were observed, and the results showed that the quadruple mutant of GmLHY displayed reduced plant height and shortened internodes. The levels of endogenous gibberellic acid (GA3) in Gmlhy1a1b2a2b was lower than in the wild type (WT), and the shortened internode phenotype could be rescued by treatment with exogenous GA3. In addition, the relative expression levels of GA metabolic pathway genes in the quadruple mutant of GmLHY were significantly decreased in comparison to the WT. These results suggest that GmLHY encodes an MYB transcription factor that affects plant height through mediating the GA pathway in soybean. We also developed genetic markers for identifying mutants for application in breeding studies. Conclusions: Our results indicate that CRISPR/Cas9-mediated targeted mutagenesis of four GmLHY genes reduces soybean plant height and shortens internodes from 20 to 35 days after emergence (DAE). These findings provide insight into the mechanisms underlying plant height regulatory networks in soybean.


2019 ◽  
Vol 19 (1) ◽  
Author(s):  
Qun Cheng ◽  
Lidong Dong ◽  
Tong Su ◽  
Tingyu Li ◽  
Zhuoran Gan ◽  
...  

Abstract Background Soybean (Glycine max) is an economically important oil and protein crop. Plant height is a key trait that significantly impacts the yield of soybean; however, research on the molecular mechanisms associated with soybean plant height is lacking. The CRISPR (clustered regularly interspaced short palindromic repeat)/Cas9 (CRISPR-associated system 9) system is a recently developed technology for gene editing that has been utilized to edit the genomes of crop plants. Results Here, we designed four gRNAs to mutate four LATE ELONGATED HYPOCOTYL(LHY) genes in soybean. In order to test whether the gRNAs could perform properly in transgenic soybean plants, we first tested the CRISPR construct in transgenic soybean hairy roots using Agrobacterium rhizogenes strain K599. Once confirmed, we performed stable soybean transformation and obtained 19 independent transgenic soybean plants. Subsequently, we obtained one T1 transgene-free homozygous quadruple mutant of GmLHY by self-crossing. The phenotypes of the T2-generation transgene-free quadruple mutant plants were observed, and the results showed that the quadruple mutant of GmLHY displayed reduced plant height and shortened internodes. The levels of endogenous gibberellic acid (GA3) in Gmlhy1a1b2a2b was lower than in the wild type (WT), and the shortened internode phenotype could be rescued by treatment with exogenous GA3. In addition, the relative expression levels of GA metabolic pathway genes in the quadruple mutant of GmLHY were significantly decreased in comparison to the WT. These results suggest that GmLHY encodes an MYB transcription factor that affects plant height through mediating the GA pathway in soybean. We also developed genetic markers for identifying mutants for application in breeding studies. Conclusions Our results indicate that CRISPR/Cas9-mediated targeted mutagenesis of four GmLHY genes reduces soybean plant height and shortens internodes from 20 to 35 days after emergence (DAE). These findings provide insight into the mechanisms underlying plant height regulatory networks in soybean.


2017 ◽  
Vol 32 (2) ◽  
pp. 135-140 ◽  
Author(s):  
M. Ryan Miller ◽  
Jason K. Norsworthy

AbstractTo address recent concerns related to auxin herbicide drift onto soybean, a study was developed to understand the susceptibility of the reproductive stage of soybean to a new auxin herbicide compared with dicamba. Florpyrauxifen-benzyl is under development as the second herbicide in a new structural class of synthetic auxins, the arylpicolinates. Field studies were conducted to (1) evaluate and compare reproductive soybean injury and yield following applications of florpyrauxifen-benzyl or dicamba across various concentrations and reproductive growth stages and (2) determine whether low-rate applications of florpyrauxifen-benzyl or dicamba to soybean in reproductive stages would have similar effect on the progeny of the affected plants. Soybean were treated with 0, 1/20, or 1/160, of the 1X rate of florpyrauxifen-benzyl (30 g ai ha−1) or dicamba (560 g ae ha−1) at R1, R2, R3, R4, or R5 growth stage. Soybean plant height and yield was reduced from 1/20X dicamba across all reproductive stages. High drift rates (1/20X) of florpyrauxifen-benzyl also reduced soybean plant height >25% and yield across R1 to R4 stages. Germination, stand, plant height, and yield of the offspring of soybean plants treated with dicamba and florpyrauxifen-benzyl were significantly affected. Dicamba applied at a rate of 1/20X at R4 and R5 resulted in 20% and 35% yield reduction for the offspring, respectively. A similar reduction occurred from florpyrauxifen-benzyl applied at R4 and R5 at the 1/20X rate, resulting in 15% to 24% yield reduction for the offspring, respectively. Based on these findings, it is suggested that growers use caution when applying these herbicides in the vicinity of reproductive soybean.


AgriPeat ◽  
2019 ◽  
Vol 19 (01) ◽  
pp. 30-36
Author(s):  
Administrator Journal

ABSTRACThis study aims to find the most potent combination of local microorganisms as a liquidbiofertilizer on marginal land, such as peatland. The study used Completely Randomized Design onpeat soil media in polybags, with 8 (eight) treatments and 4 (four) replicates, namely liquidbiofertilizer composition comprising a group of local microorganisms: 1) KHY, 2) IBT, 3) KHY +IBT, 4) KHY + IGT, 5) IBT + IGT, 6) KHY + IBT + IGT, 7) EM4 (Control +), 8) Aquadest(Controls -). The variables observed were plant height, number of leaves, number of branches,number of flowers of soybean crop. The results showed that the composition of the liquidbiofertilizer EM4 gave the best results on the parameters of plant height, the number of leaves, thenumber of branches, and the amount of flowers on soybean plants tested in peat soil media. Thecombined composition of local microorganisms KHY + IBT + IGT containing bacterial populationsof phosphate solvent, nitrogen-fixing bacteria, and cellulitic fungi, are still in the same group asEM4 in the treatment of the number of leaves and the number of plant flowers, therefore thecomposition of these potential local microorganisms is developed to become a liquid biofertilizerKeywords: Liquid Biofertilizer, Soybean Growth, Peat Land


2005 ◽  
Vol 17 (3) ◽  
pp. 291-297 ◽  
Author(s):  
André Luís Thomas ◽  
Ladaslav Sodek

Flooding leads to hypoxia, a stress to which symbiotic N2 fixation is especially sensitive. The response of fully nodulated soybean plants to a 21-day period of flooding was studied by measurements of growth parameters and xylem transport of organic nitrogenous components to the shoot, in the presence and absence of NO3- and NH4+ in the medium. Flooding was found to seriously impair N2 fixation, irrespective of the N source, as indicated by strongly reduced xylem ureide levels. In the absence of a source of N, growth was strongly reduced during flooding while accumulation of N in the shoot was virtually abolished. Flooding in the presence of 5 mM NO3- or NH4+ led to the accumulation of total N in the shoot but only NO3- promoted increases in total dry matter, plant height and leaf area above that found in the absence of N. The accumulation of N, however, was lower than that of the non-flooded control for both NO3- and NH4+. The increases in total dry matter, plant height and leaf area with NO3- was as high as those of the non-flooded control. These data clearly show the beneficial effects of NO3- during a prolonged period of flooding of the nodulated root system of soybean.


Plants ◽  
2021 ◽  
Vol 10 (8) ◽  
pp. 1737
Author(s):  
Fuxin Shan ◽  
Rui Zhang ◽  
Jin Zhang ◽  
Chang Wang ◽  
Xiaochen Lyu ◽  
...  

Excessive plant height is an important factor that can lead to lodging, which is closely related to soybean yield. Gibberellins are widely used as plant growth regulators in agricultural production. Gibberellic acid (GA3), one of the most effective active gibberellins, has been used to regulate plant height and increase yields. The mechanism through which GA3 regulates internode elongation has been extensively investigated. In 2019 and 2020, we applied GA3 to the stems, leaves, and roots of two soybean cultivars, Heinong 48 (a high-stalk cultivar) and Henong 60 (a dwarf cultivar), and GA3 was also applied to plants whose apical meristem was removed or to girded plants to compare the internode length and stem GA3 content of soybean plants under different treatments. These results suggested that the application of GA3 to the stems, leaves, and roots of soybean increased the internode length and GA3 content in the stems. Application of GA3 decreased the proportion of the pith in the soybean stems and primary xylem while increasing the proportion of secondary xylem. The apical meristem is an important site of GA3 synthesis in soybean stems and is involved in the regulation of stem elongation. GA3 was shown to be transported acropetally through the xylem and laterally between the xylem and phloem in soybean stems. We conclude that the GA3 level in stems is an important factor affecting internode elongation.


2020 ◽  
Vol 5 (1) ◽  
pp. 22-26
Author(s):  
Rahman Hairuddin

This study was aimed to find out the effective local microorganism of fruit wastes concentration on the growth and production of the soybean plants. This study was conducted in the trial land of Campus 2, Faculty of Agriculture, Cokroaminoto University, Palopo from September to December 2019. The method used in this study was the randomized block design with 5 treatments and 4 replications, therefore there were 20 experimental units. Each experimental unit comprised 2 plant units, therefore there were 40 plant samples on the given concentration of P0: Control, P1: 50 mL/L fruit waste local microorganism concentration, P2: 100 mL/L fruit waste local microorganism concentration, P3: 150 mL/L fruit waste local microorganism concentration, and P4: 200 mL/L fruit waste local microorganism concentration. The results showed that the local microorganism application on the soybean plants had no significant effects on the plant height, total of leaves, flowering period, and a total of pods. The local fruit waste microorganism with 100 mL concentration was capable of improving the plant height, total of leaves, flowering period, and a total of pods with the respective average of 31.9 cm, 51.6 leaves, 21,8 days, and 35 soybean pods


HortScience ◽  
1998 ◽  
Vol 33 (3) ◽  
pp. 449f-450
Author(s):  
Lisa M. Barry ◽  
Michael N. Dana

Legumes are grown as nurse crops in agriculture because they increase soil microbial life and productivity. Native legumes have potential in ecological restoration to mimic the benefits found in agriculture plus they enhance the restored ecosystem. This study was initiated to compare the growth rates, nodulation characteristics, and nitrogen fixation rates of a native versus a non-native legume. The two legumes were partridge pea (Cassia fasciculata); a native, wild, annual legume and soybean (Glycine max `Century Yellow); a domesticated, agricultural, annual legume native to Asia. Plants were grown for 11 weeks in pots containing silica sand and received a nitrogen-free Hoagland's nutrient solution. Beginning at week 12, plants were harvested weekly for four consecutive weeks. Nodulated root systems were exposed to acetylene gas and the resulting ethylene amounts were measured. The two legumes exhibited significant differences in nodule size and shape and plant growth rate. In soybean, nodules were large, spherical, and clustered around the taproot while in partridge pea, nodules were small, irregularly shaped, and spread throughout the fibrous root system. Soybean plants had a significantly faster growth rate at the onset of the experiment but partridge pea maintained a constant growth rate and eventually exceeded soybean plant size. In spite of these observed differences, partridge pea and soybean plants were equally efficient at reducing acetylene to ethylene. These results indicate partridge pea has the potential to produce as much nitrogen in the field as soybean. Native legumes such as partridge pea deserve further research to explore their use as nurse crops in agricultural or restoration regimes.


1981 ◽  
Vol 38 (1) ◽  
pp. 99-112
Author(s):  
Paulo R.C. Castro ◽  
Roberto S. Moraes

This research deals with the effects of growth regulators on flowering and pod formation in soybean plant (Glycine max cv. Davis). Under greenhouse conditions, soybean plants were sprayed with 2,3,5-triiodobenzoic acid (TIBA) 20 ppm, Agrostemmin (1g/10 ml/3 l) gibberellic acid (GA) 100 ppm, and (2-chloroethyl) trimethylammonium chloride (CCC) 2,000 ppm. Application of TIBA increased number of flowers. 'Davis' soybean treated with CCC and TIBA presented a tendency to produce a lower number of pods.


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