Induction of endodormancy in crown buds of leafy spurge (Euphorbia esula L.) implicates a role for ethylene and cross-talk between photoperiod and temperature.

Leafy spurge is a model for studying well-defined phases of dormancy in underground adventitious buds (UABs) of herbaceous perennial weeds, which is a primary factor facilitating their escape from conventional control measures. A 12-week ramp down in both temperature (27 â†’ 10 °C) and photoperiod (16 â†’ 8 h light) is required to induce a transition from para- to endo-dormancy in UABs of leafy spurge. To evaluate the effects of photoperiod and temperature on molecular networks of UABs during this transition, we compared global transcriptome data-sets obtained from leafy spurge exposed to a ramp down in both temperature and photoperiod (RDtp) versus a ramp down in temperature (RDt) alone. Analysis of data-sets indicated that transcript abundance for genes associated with circadian clock, photoperiodism, flowering, and hormone responses (CCA1, COP1, HY5, MAF3, MAX2) preferentially increased in endodormant UABs. Gene-set enrichment analyses also highlighted metabolic pathways involved in endodormancy induction that were associated with ethylene, auxin, flavonoids, and carbohydrate metabolism; whereas, sub-network enrichment analyses identified hubs (CCA1, CO, FRI, miR172A, EINs, DREBs) of molecular networks associated with carbohydrate metabolism, circadian clock, flowering, and stress and hormone responses. These results helped refine existing models for the transition to endodormancy in UABs of leafy spurge, which strengthened the roles of circadian clock associated genes, DREBs, COP1-HY5, carbohydrate metabolism, and involvement of hormones (ABA, ethylene, and strigolactones). We further examined the effects of ethylene by application of 1-aminocyclopropane-1-carboxylate (ACC) to paradormant plants without a ramp down treatment. New vegetative growth from UABs of ACC-treated plants resulted in a dwarfed phenotype that mimicked the growth response in RDtp-induced endodormant UABs. The results of this study provide new insights into dormancy regulation suggesting a short-photoperiod treatment provides an additive cross-talk effect with temperature signals that may impact ethylene's effect on AP2/ERF family members.

Dehydration and vernalization treatments identify overlapping molecular networks impacting endodormancy maintenance in leafy spurge crown buds.

Leafy spurge (Euphorbia esula L.) is a herbaceous perennial weed that reproduces vegetatively from an abundance of underground adventitious buds (UABs), which undergo well-defined phases of seasonal dormancy (para-, endo-, and ecodormancy). In this study, the effects of dehydration stress on vegetative growth and flowering potential from endodormant UABs of leafy spurge was monitored. Further, microarray analysis was used to identify critical signaling pathways of transcriptome profiles associated with endodormancy maintenance in UABs. Surprisingly, only 3-day of dehydration stress is required to break the endodormant phase in UABs; however, the dehydration-stress treatment did not induce flowering. Previous studies have shown that prolonged cold treatment of UABs breaks endodormancy and induces a vernalization response leading to flowering. Thus, in this study, comparing transcriptome data from UABs exposed to short-term dehydration and vernalization provided a unique approach to identify overlapping molecular mechanisms involved in endodormancy maintenance and floral competence. Analysis of transcriptome data associated with breaking endodormancy by both environmental treatments identified LEC1, PHOTOSYSTEM I RC, and brassinosteroids as common central hubs of upregulated genes, while DREB1A, CBF2, GPA1, MYC2, bHLH, BZIP, and flavonoids were identified as common central hubs of downregulated genes. The majority of over-represented gene sets common to breaking endodormancy by dehydration stress and vernalization were downregulated and included pathways involved in hormone signaling, chromatin modification, and circadian rhythm. Additionally, the over-represented gene sets highlighted pathways involved in starch and sugar degradation and biogenesis of carbon skeletons, suggesting a high metabolic activity is necessary during the endodormant phase. The data presented in this study helped to refine our previous model for dormancy regulation.

Low temperatures impact dormancy status, flowering competence, and transcript profiles in crown buds of leafy spurge.

Leafy spurge (Euphorbia esula) is an herbaceous perennial weed that produces vegetatively from an abundance of underground adventitious buds. In this study, we report the effects of different environmental conditions on vegetative production and flowering competence, and determine molecular mechanisms associated with dormancy transitions under controlled conditions. Reduction in temperature (27-10 degrees C) and photoperiod (16-8 h) over a 3-month period induced a para- to endo-dormant transition in crown buds. An additional 11 weeks of cold (5-7 degrees C) and short-photoperiod resulted in accelerated shoot growth from crown buds, and 99% floral competence when plants were returned to growth-promoting conditions. Exposure of paradormant plants to short-photoperiod and prolonged cold treatment alone had minimal affect on growth potential and resulted in ~1% flowering. Likewise, endodormant crown buds without prolonged cold treatment displayed delayed shoot growth and ~2% flowering when returned to growth-promoting conditions. Transcriptome analysis revealed that 373 and 260 genes were differentially expressed (P < 0.005) during para- to endo-dormant and endo- to eco-dormant transitions, respectively. Transcripts from flower competent vs. non-flower competent crown buds identified 607 differentially expressed genes. Further, sub-network analysis identified expression targets and binding partners associated with circadian clock, dehydration/cold signaling, phosphorylation cascades, and response to abscisic acid, ethylene, gibberellic acid, and jasmonic acid, suggesting these central regulators affect well-defined phases of dormancy and flowering. Potential genetic pathways associated with these dormancy transitions and flowering were used to develop a proposed conceptual model.

A psbA mutation in Kochia scoparia (L) Schrad from railroad rights-of-way with resistance to diuron, tebuthiuron and metribuzin.

Kochia [Kochia scoparia (L) Schrad] has become resistant to many herbicides used in cropland and railroad rights-of-way in North Dakota and Minnesota. Kochia scoparia plants that had survived annual treatments with diuron and tebuthiuron were sampled along railroad rights-of-way in North Dakota and Minnesota. The samples were screened in the greenhouse for resistance to diuron, tebuthiuron, metribuzin and bromoxynil from 0.5x to 32x the recommended use rates. A resistant K scoparia accession (MN-3R) was confirmed with resistance up to 16-fold higher than recommended use rates for tebuthiuron and diuron and up to 4-fold higher for metribuzin. However, the resistant K scoparia accession was susceptible to bromoxynil even at 50% of the recommended use rate. The herbicide binding region of the psbA gene fragment of eight resistant (R) and seven susceptible (S) K scoparia accessions was PCR-amplified and sequenced for detection of mutations. The psbA gene of four R K scoparia accessions was mutated at residue 219 with substitution of isoleucine for valine (GenBank accession number AY251265). The seven S K scoparia accession sequences were wild-type at this residue (GenBank accession number AY251266). The other four R accessions sequences showed a previously known triazine R mutation with substitution of glycine for serine at residue 264. All 15 K scoparia accessions were wild-type at all other psbA residues within the region analyzed. Resistance to diuron, tebuthiuron and metribuzin among the railroad rights-of-way K scoparia is probably due to the mutation at residue 219 of the psbA gene in some plants, but due to the previously reported Ser(264)Gly substitution in other plants. Target-site resistance associated with a change of valine to isoleucine at residue 219 of the psbA target-site in weeds has previously been reported for Poa annua L selected in diuron-treated grass seed fields, and for Amaranthus powelli S Wats selected in linuron-treated carrot fields. This is the first report of the mutation in herbicide-resistant K scoparia.

An isoleucine to leucine mutation in acetyl-CoA carboxylase confers herbicide resistance in wild oat.

Wild oat (Avena fatua L.) populations resistant to herbicides that inhibit acetyl-CoA carboxylase (ACCase; EC 6.4.1.2) represent an increasingly important weed control problem. The objective of this study was to determine the ACCase mutation responsible for herbicide resistance in a well-studied wild oat biotype (UMI). A 2039-bp region encompassing the carboxybiotin and acetyl-CoA binding domains of multifunctional plastidic ACCase was analyzed. DNA sequences representing three plastidic ACCase gene loci were isolated from both the resistant UMI and a herbicide-susceptible biotype, consistent with the hexaploid nature of wild oat. Only one nonsynonymous point mutation was found among the resistant wild oat sequences, inferring an isoleucine to leucine substitution. The position of this substitution corresponds to residue 1769 of wheat (Triticum aestivum L.) plastidic ACCase (GenBank accession No. AF029895). Analysis of an F2 population derived from a cross between a herbicide-resistant and a susceptible biotype confirmed co-segregation of herbicide resistance with the mutated ACCase. We conclude that the isoleucine to leucine mutation is responsible for herbicide resistance in UMI wild oat based on a comparison of the substitution site across species and ACCase types. While isoleucine is conserved among plastidic ACCases of herbicide-susceptible grasses, leucine is found in plastidic and cytosolic forms of multifunctional herbicide-resistant ACCase.