Effect of different herbicides on Bidens pilosa and Euphorbia heterophylla biotypes resistant to ALS inhibitors

Select herbicides with different mechanism of action is a satisfactory option for resistant weed control. Then, the present work aimed to study the efficiency of different herbicides and their mixtures on Bidens pilosa (blackjack) and Euphorbia heterophylla (wild poinsettia) biotypes, resistant to ALS herbicides in two development stages. The trials we arranged in a completely randomized design with four replications. The treatments tested were (g a.i/a.e ha-1): imazethapyr at 70 and 140 (WG formulation) + 1.0% Assist; imazethapyr at 57.6 and 72 (SL formulation) + 1.0% Assist; imazapic + imazethapyr at 56 and 70 + 1.0% Assist; glyphosate + imazethapyr (596); saflufenacil + glyphosate at 35 + 720 + 0.5% Dash in tank mix, glyphosate at 720 and, a control without herbicide application. Control efficiency was evaluated, as well as dry matter accumulation at the end of the studies. Plants of both species were more susceptible to herbicides at the early stage of development (2 to 4 leaves). The treatments with saflufenacil + glyphosate, (imazethapyr + glyphosate) and glyphosate promoted the best controls, regardless of the species studied and the application stage. The mixture with saflufenacil provided the highest control speed, and the mixture (imazethapyr + glyphosate) was less efficient among three excellent treatments when applied to plants in the 4-6 leaf stage. The treatments (imazethapyr, in both formulations) and (imazethapyr + imazapic) were ineffective in controlling the studied biotypes, regardless of dose and developmental stage studied.

Characterization of the acetohydroxyacid synthase multigene family in the tetraploide plant Chenopodium quinoa

Background Currently, the technology called Clearfield® is used in the development of crops resistant to herbicides that inhibit the enzyme acetohydroxy acid synthase (AHAS, EC 2.2.1.6). AHAS is the first enzyme of the biosynthetic pathway that produces the branched-chain of the essential amino acids valine, leucine, and isoleucine. Therefore, multiple copies of the AHAS gene might be of interest for breeding programs targeting herbicide resistance. In this work, the characterization of the AHAS gene was accomplished for the Chenopodium quinoa Regalona-Baer cultivar. Cloning, sequencing, and Southern blotting were conducted to determine the number of gene copies. Results The presence of multiple copies of the AHAS gene as has been shown previously in several other species is described. Six copies of the AHAS gene were confirmed with Southern blot analyses. CqHAS1 and CqAHAS2 variants showed the highest homology with AHAS mRNA sequences found in the NR Database. A third copy, CqAHAS3, shared similar fragments with both CqAHAS1 and CqAHAS2, suggesting duplication through homeologous chromosomes pairing. Conclusions The presence of multiple copies of the gene AHAS shows that gene duplication is a common feature in polyploid species during evolution. In addition, to our knowledge, this is the first report of the interaction of sub-genomes in quinoa.

Effects of knockout of 2,3-butanediol synthesis key enzyme genes on 1,3-propandediol production in Klebsiella pneumoniae / 生物工程学报

2,3-butanediol (2,3-BD) is a major byproduct of 1,3-propandediol (1,3-PDO) fermentation by Klebsiella pneumoniae. To decrease the formation of 2,3-BD, the budC and budA gene, coding two key enzymes of 2,3-BD synthetic pathway in K. pneumoniae, were knocked out using Red recombination technology. The growth of the two mutants were suppressed in different level. The budC deficient strain fermentation results showed that 1,3-PDO concentration increased to 110% and 2,3-butanediol concentration dropped to 70% of the parent strain. However, the budA deficient strain did not produce 1,3-PDO and 2,3-BD, and the final titer of lactic acid, succinic acid, ethanol and acetic acid increased remarkably compared with the parent strain. Further analysis of budC deficient strain fermentation inferred that K. pneumoniae possessed the 2,3-BD cycle as a replenishment pathway. The consequence provided a new evidence for reforming low-byproduct K. pneumoniae.

Efeito do déficit hídrico na eficiência de herbicidas e nas características bioquímicas de picão-preto / Effect of water deficit on the efficiency of herbicides and biochemical characteristics of hair beggarticks

Este estudo teve como objetivo avaliar a eficiência de herbicidas inibidores da acetolactato sintase (ALS) e protoporfirinogênio oxidase (PROTOX) no controle de Bidens pilosa sob duas condições hídricas, bem como determinar a influência deste déficit hídrico sob o conteúdo de carboidratos e proteínas solúveis da planta daninha. O delineamento experimental utilizado foi inteiramente casualizado com os tratamentos, com quatro repetições, dispostos em um esquema fatorial 4x2, sendo quatro herbicidas (fomesafen, lactofen, chlorimuron-ethyl e imazethapyr, na dose recomendada pelo fabricante) e duas condições hídricas (-0,5 MPa e -0,01 MPa). Aos 7, 14, 21 e 28 dias após a aplicação (DAA) dos tratamentos foi avaliada de forma visual a eficiência de controle dos herbicidas. Para a determinação dos solutos orgânicos foram coletadas plantas às 24, 48, 72 e 96 horas após a aplicação (HAA). Os herbicidas controlaram de forma eficiente as plantas de B. pilosa, com exceção do lactofen, enquanto que o déficit hídrico afetou a eficiência de controle de todos os herbicidas testados. O conteúdo de carboidratos solúveis aumentou com a imposição do déficit hídrico, contudo, os herbicidas e o déficit hídrico proporcionaram redução de proteínas solúveis nas plantas de B. pilosa.

The objective of this work was to evaluate conditions the effectiveness of acetolactate synthase (ALS) and protoporphyrinogen oxidase (PROTOX) inhibitors in the Bidens pilosa control under two water deficit conditions, as well as to determine the influence of water deficit under the contents of soluble carbohydrates and protein of weed. The experimental design was randomized completely design with the treatments, with four replications, setup in a factorial scheme 4x2, with four herbicides (fomesafen lactofen, chlorimuron-ethyl and imazethapyr), and two water conditions (-0.5 MPa and -0.01MPa). At 7, 14, 21 and 28 days after application (DAA), was assessed visually control efficiency of herbicides. For the determination of organic solutes plants were collected at 24, 48, 72 and 96 hours after application (HAA). Herbicides controlled efficiently plants of B. pilosa, except lactofen, whereas the water deficit affected the efficiency of control of all the herbicides. The soluble carbohydrate content increased with the imposition of water deficit, however, herbicides and water deficit further reduction of soluble proteins in plants of B. pilosa.

Comparative study of Chalcone synthase promoters across plant families / Estudio Comparativo de Promotores de la Chalcón Sintasa en Diferentes Familias de Plantas

In the post – genomic era the understanding of gene regulation has become a challenge and a research priority. In this research, we performed a comparative study of the regulator sequences of the chalcone synthase gene across plant families. Twenty-two sequences of chalcone synthase promoters were compared considering three regulator Cis elements: G-Box, H-Box and TATA Box. Our results show that these Cis elements are conserved among species and even at the family level. However, in some species all of the Cis elements were not found, showing that the expression and regulation of these promoters via the Cis elements can be variable. Additionally, a comparison between promoters from a species with a chalcone synthase multigene family showed that the duplicate genes are variable in the composition of the Cis elements, suggesting that these genes could be expressing in different ways.

Expression of acetohydroxyacid synthase isozyme genes ilvBN, ilvGM, ilvIH and their resistance to AHAS-inhibitor herbicides / 生物工程学报

Acetohydroxyacid synthase (AHAS) catalyses the first reaction in the pathway for synthesis of the branched-chain amino acids. AHAS is the target for sulfonylurea, imidazolinone and other AHAS-inhibitor herbicides. Herbicides-resistant AHAS genes have potential application in plant transgenetic engineering and development of new generation herbicide. The AHAS isozyme genes ilvBN, ilvGM and ilvIH were cloned from metsulfuron-methyl resistant strain Klebsiella sp. HR11 and metsulfuron-methyl sensitive strain Klebsiella pneumoniae MGH 78578. Homologous sequences comparison indicated that the differences in AHAS isozyme genes at amino acid levels between strain HR11 and strain MGH 78578 were mainly on the large subunits of ilvBN and ilvGM. The three AHAS isozyme genes from HR11 and MGH 78578 were ligated into the expression vector pET29a(+) and expressed in Escherichia coli BL21, respectively. The results of enzyme inhibition assay showed that only ilvBN and ilvGM from strain HR11 showed strong resistance to AHAS-inhibitor herbicides, while ilvIH from strain HR11 and ilvBN, ilvGM and ilvIH from strain MGH78578 were sensitive to AHAS-inhibitor herbicides.

Eucalyptus ESTs associated with resistance to herbicide inhibitors of aromatic and branched-chain amino acid synthesis

Herbicides inhibit enzymatic systems of plants. Acetolactate synthase (ALS, EC = 4.1.3.18) and 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS, EC 2.5.1.19) are key enzymes for herbicide action. Hundreds of compounds inhibit ALS. This enzyme is highly variable, enabling the selective control of weeds in a number of crops. Glyphosate, the only commercial herbicide inhibiting EPSPS is widely used for non-selective control of weeds in many crops. Recently, transgenic crops resistant to glyphosate were developed and have been used by farmers. The aim of this study was the data mining of eucalypt expressed sequence tags (ESTs) in the FORESTs Genome Project database (https://forests.esalq.usp.br) related to these enzymes. Representative amino acid sequences from the NCBI database associated with ALS and EPSPS were blasted with ESTs from the FORESTs database using the tBLASTx option of the blast tool. The best blasting reads and clusters from FORESTs, represented as nucleotide sequences, were blasted back with the NCBI database to evaluate the level of similarity with available sequences from different species. One and seven clusters were identified as showing high similarity with EPSPS and ALS sequences from the literature, respectively. The alignment of EPSPS sequences allowed the identification of conserved regions that can be used to design specific primers for additional sequencings

Genetically modified industrial brewing yeast with high-glutathione and low-diacetyl production / 生物工程学报

Recombinant plasmid pICG was constructed by replacing the internal fragment of a-acetohydroxyacid synthase (AHAS) gene (ILV2) with a copy of gamma-glutamylcysteine synthetase gene (GSH1) and copper chelatin gene (CUP1) from the industrial brewing yeast strain YSF31. YSF31 was transformed with plasmid pICG linearized by Kpn I and Pst I. A recombinant strain with high-glutathione and low-diacetyl production was selected. The results of fermentation in 100-L bioreactor showed that the lagering time of beer produced for recombinant strain T2 was shortened by 3 days and the shelf life of the beer was prolonged about 50%. It may be more acceptable for the commercial application, as it does not contain foreign DNA.