Native instruments komplete 9 ultimate d14 megashare

ccd, carotenoid cleavage dioxygenase d14, dwarf 14 EIC, extracted ion chromatogram EV, empty vector Kcps, kilo counts per second max2, more axillary growth 2 MRM, multiple reaction monitoring SL, strigolactone UHPLC, ultra-high-performance liquid chromatography. Values for graphs (B) and (F) are listed in S1 Data. Three independent experiments were performed at different developmental stages (two-tailed Student t test). Quantifier ion (m/z) 347.1 > 233.1 was selected to achieve maximum signal intensity. (F) (В±)-2вЂІ- epi-orobanchol levels (Kcps) in root exudates of hydroponic EV, max2, d14, and ccd7 plants (В±SE, n = 3вЂ“4) were analyzed by targeted UHPLC-triple quadrupole-MS metabolomics. (E) Representative chromatograms of (В±)-2вЂІ- epi-orobanchol in root exudates, as analyzed by targeted UHPLC-triple quadrupole-MS metabolomics different MRM ions were listed on the top of the panel. (D) Comparison of fragmentation patterns of root exudates and (В±)-2вЂІ- epi-orobanchol by high-resolution tandem MS. (C) Representative chromatograms of root exudates, (В±)-2вЂІ- epi-orobanchol, (В±) orobanchol, and (В±) strigol analyzed under positive ionization mode. (B) Relative transcript abundance of CCD7 and CCD8 in roots of EV, d14, and max2 plants (В±SE, n = 3вЂ“4). (A) A scheme of SL biosynthesis and perception in N. S3 Fig: Orobanchol levels in root exudates of max2, d14, and ccd7 plants. hybrida Ps, Pisum sativum Pt, Populus trichocarpa Sl, S. thaliana CCD, carotenoid cleavage dioxygenase d14, dwarf14 max2, more axillary growth 2 Mt, Medicago truncatula Na, N. Values for graphs (C-E) are listed in S1 Data. Primers for constructing vectors are described in S2 Table. (E) Vector map of pSOL8CCD7 transformation construct used for constructing ccd7 lines and relative signal intensity of ccd7#1 and ccd7#2 plants with indicated primers. Relative signal intensity of d14#1 and d14#2 (C) and kai2#1 and kai2#2 (D) plants was quantified with the indicated primers. The basic architecture of the vector was the same as shown for the vector described in S1D Fig with d14 and kai2 fragments replacing the concatenated max2 fragments. (C and D) pRESC8D14 and pRESC8KAI2 were used for the construction of the d14 and kai2 lines. (A and B) Phylogenetic analysis of D14 and KAI2 (A), CCD7 and CCD8 (B) gene families from At, Os, Sl, Na, Ph, Ps, Mt, and Pt. S2 Fig: Characterization of D14, CCD7, and KAI2 genes in N. AD, activation domain ago 8, argonaute 8 BD, binding domain CA, Capsicum annuum D14, dwarf14 eGFP, enhanced green fluorescent protein EV, empty vector KAR, karrikin LB, left border MAX2, more axillary growth 2 Na, N. Values for graphs (D-G) are listed in S1 Data. (G) Seed germination ratios of EV, max2#1, and max2#2 seeds with KAR1 (0.01 ng/ОјL) treatment for 9 days (В±SE, 4 replicates, each replicates included 25 seeds) (* P < 0.05 ** P < 0.01 *** P < 0.001 two-tailed Student t test). Hypocotyl length of EV, max2#1, and max2#2 seedlings without treatment and relative hypocotyl length of EV, max2#1, and max2#2 seedlings with indicated treatment (В±SE, n = 11вЂ“15). (F) Representative hypocotyl phenotypes of EV, max2#1, and max2#2 seedlings after growth with 5 ОјM rac-GR24 or 5 ОјM KAR1 treatment for 7 days. pVS1_3 and nptII_1 (red) are probes indicating over-reads of the plasmid sequences outside of the T-DNA borders, which do not affect the plant phenotype or its stable inheritance note that the lack of signal from these probes reveals that over-reads did not occur during the transformation process. Briefly, TNOS+LB_1, hptII_3, PNOS_1, P35S_1, and T35S_1 (blue) are used to evaluate single and complete T-DNA insertions. The single and complete T-DNA insertion in both max2#1 and max2#2 plants was validated by using NanoStrings nCounter technology as described in. A published single-copy insertion of the stably transformed line ( ago8) was used as a positive control.

(E) Relative signal intensity of max2#1 and max2#2 plants with indicated primers. Primers designed within the LB and RB region (blue) and outside the LB and RB region (red) were used in (F). (D) Vector map of pRESC8MAX2 transformation construct. The transformants were grown on QDO (SD в€’Ade/в€’His/в€’Leu/в€’Trp). GAL4 DNA-BD-NaD14 and AD-NaMAX2a/2b were cotransformed into yeast. (C) Interactions between NaMAX2a/b and NaD14 proteins in the presence of rac-GR24 as evaluated by yeast two-hybrid assays. benthamiana leaves 72 hours after inoculation. MAX2a-eGFP and MAX2b-eGFP were transiently expressed in N.

(B) Subcellular localization of MAX2a and MAX2b. (A) Phylogenetic analysis of MAX2 gene family from CA, Na, Ph, Sl, and St. S1 Fig: Characterization of MAX2 gene in N.

Native instruments komplete 9 ultimate d14 megashare