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Purpose: Purple acid phosphatases (PAPs) includ the largest classes of non-specific plant acid phosphatases. Most characterized PAPs were found to play physiological functions in phosphorus metabolism. In this study, we investigated the function of AtPAP17 gene encoding an important purple acid phosphatase in Arabidopsis thaliana. Methods: The full-length cDNA sequence of AtPAP17 gene under the control of CaMV-35S promoter was transferred to the A. thaliana WT plant. The generated homozygote AtPAP17-overexpressed plants were compared by the types of analyses with corresponding homozygote atpap17-mutant plant and WT in both + P (1.2 mM) and − P (0 mM) conditions. Results: In the + P condition, the highest and the lowest amount of Pi was observed in AtPAP17-overexpressed plants and atpap17-mutant plants by 111% increase and 38% decrease compared with the WT plants, respectively. Furthermore, under the same condition, APase activity of AtPAP17-overexpressed plants increased by 24% compared to the WT. Inversely, atpap17-mutant plant represented a 71% fall compared to WT plants. The comparison of fresh weight and dry weight in the studied plants showed that the highest and the lowest amount of absorbed water belonged to OE plants (with 38 and 12 mg plant−1) and Mu plants (with 22 and 7 mg plant−1) in + P and − P conditions, respectively. Conclusion: The lack of AtPAP17 gene in the A. thaliana genome led to a remarkable reduction in the development of root biomass. Thus, AtPAP17 could have an important role in the root but not shoot developmental and structural programming. Consequently, this function enables them to absorb more water and eventually associated with more phosphate absorption. [ABSTRACT FROM AUTHOR]

Peanut (Arachis hypogaea L.) is an oil and economic crop of vital importance, and peanut pod is the key organ influencing the yield and processing quality. Hence, the Pod-related traits (PRTs) are considered as important agronomic traits in peanut breeding. To broaden the variability of PRTs in current peanut germplasms, three elite peanut cultivars were used to construct Ethyl methane sulfonate (EMS)-induced mutant libraries in this study. The optimal EMS treatment conditions for the three peanut varieties were determined. It was found that the median lethal dose (LD50) of EMS treatment varied greatly among different genotypes. Finally, the EMS-induced peanut mutant libraries were constructed and a total of 124 mutant lines for PRTs were identified and evaluated. Furthermore, "M-8070", one of the mutant lines for pod constriction, was re-sequenced via high-throughput sequencing technology. The genome-wide variations between "M-8070" and its wild parent "Fuhua 8" (FH 8) were detected. 2994 EMS-induced single nucleotide polymorphisms (SNPs) and 1188 insertion-deletions (InDels) between "M-8070" and its wild parent were identified. The predominant SNP mutation type was C/G to T/A transitions, while the predominant InDel mutation type was "1-bp". We analyzed the distribution of identified mutations and annotated their functions. Most of the mutations (91.68% of the SNPs and 77.69% of the InDels) were located in the intergenic region. 72 SNPs were identified in the exonic region, leading to 27 synonymous, 43 nonsynonymous and 2 stop-gain variation for gene structure. 13 Indels were identified in the exonic region, leading to 4 frame-shift, 8 non-frame-shift and 1 stop-gain variations of genes. These mutations may lead to the phenotypic variation of "M-8070". Our study provided valuable resources for peanut improvement and functional genomic research. [ABSTRACT FROM AUTHOR]

In this study, the mutants (P, 13, 17, 87, 89), 'TN57' and 'Hong Xin Wei' of sweet potato (Ipomoea batatas) were used as experimental materials. The salt stress was conducted in three aspects: tissue culture, potted trial and field experiment. The physiological and phenotype responses of sweet potato under salt stress were analyzed to evaluate the salt tolerance of the mutants. The results showed that different mutants responded differently to salt stress, and their salt tolerance index (STI) varied with different traits and decreased with increasing salt concentrations. The seedlings of tissue culture for mutants performed stably at 0.5% and was significantly inhibited at 1.0% salt concentration. Among them, the root length of No. 13 and No. 89 of mutants performed best. In addition, the results of the potted experiment showed that the root weight of sweet potato was stable under 150 mM salt stress, and the STI of the mutagenic line, No. 87, was the highest. It was worth noting that under 150 mM salt stress, sweet potato accumulated large amounts of Na+ and maltose content in the roots, which might be the reason to explain why the mutants had strong salt tolerance. Further evaluation in the results of the field experiment showed that the salt stress had a slight negative impact on the shoots of sweet potato, but had larger negative effect on the roots, and it caused to the decreasing of yield for sweet potato. However, the degree of yield reduction varied with different genotypes. Among them, 'Hong Xin Wei', P and No. 87 were significantly higher. In the potted trial and the saline soil field experiment, the sweet potato mutant P and No. 87 could grow normally and had higher yields throughout the whole cultivation period. It indicated that the two lines had higher salt tolerance and could be used as the experimental materials of salt-tolerance in the breeding program of sweet potato. [ABSTRACT FROM AUTHOR]

Novel mutant alleles related to isoflavone content are useful for breeding programs to improve the disease resistance and nutritional content of soybean. However, identification of mutant alleles from high-density mutant libraries is expensive and time-consuming because soybean has a large, complicated genome. Here, we identified the gene responsible for increased genistein-to-daidzein ratio in seed of the mutant line F333ES017D9. For this purpose, we used a time- and cost-effective approach based on selective genotyping of a small number of F2 plants showing the mutant phenotype with nearest-neighboring-nucleotide substitution- high-resolution melting analysis markers, followed by alignment of short reads obtained by nextgeneration sequencing analysis with the identified locus. In the mutant line, GmCHR5 harbored a single-base deletion that caused a change in the substrate flow in the isoflavone biosynthetic pathway towards genistein. Mutated GmCHR5 was expressed at a lower level during seed development than wild-type GmCHR5. Ectopic overexpression of GmCHR5 increased the production of daidzein derivatives in both the wild-type and mutant plants. The present strategy will be useful for accelerating identification of mutant alleles responsible for traits of interest in agronomically important crops. [ABSTRACT FROM AUTHOR]

A protocol for generating aluminium (Al) tolerant sugarcane using 5-azacytidine (5-azaC)-induced epimutagenesis was developed. Eight (8) plantlets per gram of cultivar N51 calli were generated from the 5-azaC (100 μM) and Al (1.5–50 mM) treatment (+Az + Al) when compared with 104 plantlets from non-mutagenised, no Al control treatment. Following in vitro selection on 60 mM Al, ex vitro stress over four rounds of chimera dissolution during vegetative propagation was applied. In the final stress round, 36% of the Mut 2 epilines survived with plants showing higher relative water content (61.2%) when compared with 47.3% from the stressed control (S N51). The Mut 2 line maintained a higher green leaf area (83.4%) and longer roots (32.4 cm) under stress than S N51 (61.4% and 26.3 cm, respectively). Overall, Mut 2 had a high stress tolerance index of 85.4%, compared with 79.0% from the S N51, nearing that of the non-stressed N51 control (NS N51, 100%) when data were analysed using PCA and clustering analyses of morpho-physiological traits. Analysis of molecular variance (AMOVA) revealed high epigenetic differentiation (ɸst = 0.67) and a variation of 66.6% observed among N51 genotypes. The principal coordinate analysis (PCoA) showed that Mut 2 was epigenetically distinct from S N51. These findings support previous studies that 5-azaC can be used for novel trait creation via epimutagenesis and highlights the necessity for chimera dissolution to achieve stable traits in epibreeding of sugarcane. • Mut 2, had a high Al stress tolerance index of 85.4% vs 79.0% observed in the stressed control. • Epiline Mut 2 had higher relative water content (61.2%), leaf area (83.4%), and root length (32.4 cm) than stressed control (47.3%, 61.4%, 26.3 cm respectively). • High epigenetic differentiation (66.6%) between Mut 2 and control lines was recorded, indicating 5-azaC-induced epigenetic changes and distinctness. • 5-azaC enables novel trait creation and the study highlights the importance of chimera dissolution for stable traits in epibreeding. [ABSTRACT FROM AUTHOR]

In general, barley with smoother degradation of seed storage material (modification) is known to have lower seed dormancy and a higher risk of pre-harvest sprouting (PHS). The Qsd1 and Qsd2 genes affecting seed dormancy have also been reported to affect malting characteristics. To overcome this situation, barley with an excellent balance of modification and seed dormancy was screened among the mutant population derived from 'Nittakei 68 (N68)'. In the mutant lines screened, 'N68-411' exhibited a moderate seed dormancy similar to that of 'N68', but with higher modification. It was considered that 'N68-411' was mutated in a different gene from Qsd1 and Qsd2 , resulting in no significant change in seed dormancy, only in modification. 'N68-411' also showed rapid modification during germination, suggesting a possibility of shorter germination time in malting process. Analysis of progenies from 12WY375/N68-411 and Xena/N68-411 indicated that the 'N68-411' trait was inherited and associated with a small number of genes. [Display omitted] • Sustainability of barley and malt production. • Discovery of the barley mutant line with novel traits. • Better balance between malt modification and seed dormancy. • Possibility of a shorter germination time in the malting process. [ABSTRACT FROM AUTHOR]

Hybrid Rada was developed with simple cross of Bulgarian line cms2607 and mutant line 12002 R. Mutant line was developed after treatment of immature sunflower ( Helianthus annuus L.) zygotic embryos of fertility restorer line R 2574 with gamma irradiation at dose of 8 Gy for 1 min. Hybrid Rada was tested for three years in testing plots of Dobroudja Agricultural Institute - General Toshevo and for two years at the seven locations of State Variety Testing Commission. The ANOVA procedure proves that the parents (2607 A and 12002 R) and the received hybrid Rada are with different genetic potential in the studied indices. In our study, all six characters recorded positive and significant heterosis in the direction of both relative to parental average (h1) and relative to better parent (h2). The correlations of the hybrid Rada with the mean values of the parental lines (h1) and with the mean value of the parent with higher indices (h2) were statistically significant. In h1, the variation was from 1.52% to 17.92%; in h2, the variation was within 1.49-12.81%. The highest positive heterotic effect (h1 = 17.92%) in the hybrid Rada was for trait seed yield per head relative to parental average h1. The model of inheritance of studied indices is additive dominant with partial or full dominance in relation to parent with the higher value. This was markedly expressed for indices seed yield per head. [ABSTRACT FROM AUTHOR]

Rice is a source of food supply as the staple food. The narrow genetic diversity of Korean Japonica cultivars is prone to be susceptible to abiotic and biotic stresses such as rice blast, bacterial blight, and brown planthopper (BPH). The best approach for prevention of the breeding and extermination of disease and insect pest would be the development of rice cultivars harboring multiple disease and insect pest resistance genes. National Institute of Crop Science (NICS) in Korea developed a mutant line, Namil(EMS)-bl10,bph1, by using ethyl-methane-sulfonate (EMS) as the chemical mutagen on the wild type, Namil. The mutant line exhibits strong resistance levels against rice blast and brown planthopper. To localize favorable mutated allele types, two mapping populations were constructed by using F2 progenies derives crosses between Namil(EMS)-bl10,bph1 and Milyang23 and Namil (wild type) and Milyang23. Each F2 progenies were evaluated in terms of DNA marker genotype as well as agronomic traits measured, and for rice blast and BPH resistance, virulent blast isolate 94-254 and Korean biotype of BPH were used to screen F2:3 seedlings to conduct association analysis between marker genotype and evaluated phenotype of progeny lines. Through comparisons on major loci between two mapping populations, the putative major loci for rice blast resistance (RM1337, R2=54.4%) and BPH (RM0277, R2=55.1%) were primarily localized on chromosome 12. [ABSTRACT FROM AUTHOR]

Abstract: Sunflower mutant lines with an enhanced tolerance and metal accumulation capacity obtained by mutation breeding have been proposed for Zn, Cd and Cu removal from metal-contaminated soils in previous studies. However, soils contaminated with trace elements induce various biochemical alterations in plants leading to oxidative stress. There is a lack of knowledge concerning the metal accumulation and antioxidant responses during the growth and development of sunflowers. This study, therefore, aimed to characterise metal accumulation and possible metal detoxification mechanisms in young seedlings and adult sunflowers. Beside the inbred line, two mutant lines with an improved growth and enhanced metal uptake capacity on a metal contaminated soil were investigated in more detail. Sunflowers cultivated on a metal-contaminated soil in the greenhouse showed a decrease in shoot biomass and chlorophyll concentration in two different developmental stages. Adult sunflowers showed a lower sensitivity to metal toxicity than young seedlings, whereas mutant lines were more tolerant to metal stress than the control. Mutant lines also produced a higher amount of carotenoids on a metal-contaminated soil than on the control soil, indicating a possible protective mechanism of sunflower mutants against oxidative stress caused by Cd and excess Zn. Heavy metals primarily increased the activity of antioxidant enzymes involved in the ascorbate–glutathione cycle in sunflower leaves. Activity of dehydroascorbate reductase (DHAR), monodehydroascorbate reductase (MDHAR) and glutathione reductase (GR) was strongly increased in young seedlings exposed to heavy metals. The enzyme activities were even more pronounced in mutant lines. A significantly increased ascorbate peroxidase (APOX) activity in adult sunflowers exposed to heavy metals indicated an elevated use of ascorbate after a longer exposure to metal stress. An increased antioxidant level corresponded to a high Cd and Zn accumulation in young and adult sunflowers. Metal distribution, zinc translocation in particular, from the root into the shoot tissue obviously increased during sunflower growth and ripening. Altogether, these results suggest that sunflower plants, primarily the mutant lines, possess an efficient defence mechanism against oxidative stress caused by metal toxicity. A good tolerance of sunflowers toward heavy metals coupled with an increased metal accumulation capacity might contribute to an efficient removal of heavy metals from a polluted area. [Copyright &y& Elsevier]

Immature sunflower ( Helianthus annuus L.) zygotic embryos of sunflower fertility restorer line R 2574 were treated with ultrasound before planting on the embryo culture medium. As a result some chlorophyll, leaf and inflorescence mutations were observed. New sunflower forms with inherited morphological, biochemical and phytopatological changes were obtained through selection and self-pollination. The line R 12003, subjected to investigation in this study, was characterized with significant changes concerning most of the studied characters. The contribution of the present research connected to the investigated mutation was the appearance of resistance to the parasite broomrape. The increased number of seeds per head in line R 12003, increased oil content in seed and 100% resistance to the parasite Orobanche are the desired combination in the breeding programme of sunflower. [ABSTRACT FROM AUTHOR]

Hybrid Yana was developed with simple cross of Bulgarian line cms2607 and mutant line R12003. Mutant line was produced after treatment of immature sunflower ( Helianthus annuus L.) zygotic embryos of fertility restorer line R 2574 with ultrasound. In 2004, 2006 and 2007 years hybrid Yana was presented for testing at State Variety Testing Commission. The hybrid considerably exceeded the mean standard (the Bulgarian commercial hybrid Albena and French commercial hybrid Diabolo) by seed yield. The seed yield of hybrid Yana was significantly higher with 31 kg/dka, or 9.8%. The correlations of the cross cms2607 × R12003 with the mean values of the parental lines (h1) and with the mean value of the parent with higher indices (h2) were statistically significant. In h1 the variation was from 152.4% to 1471.2%; in h2 the variation was within 149.5-1052.9%. The evaluation of the heterosis effect parameters proved that the parent with the higher values of the indices was superdominant. This was markedly expressed for indices total seeds number per head and seed yield per head. Hybrid Yana possessing immunity to the Orobanche cumana population of races A-F and immunity to Plasmopara helianhi-races 300, 700 and 731. [ABSTRACT FROM AUTHOR]

Sea urchins are premier model organisms for the study of early development. However, the lengthy generation times of commonly used species have precluded application of stable genetic approaches. Here, we use the painted sea urchin Lytechinus pictus to address this limitation and to generate a homozygous mutant sea urchin line. L. pictus has one of the shortest generation times of any currently used sea urchin. We leveraged this advantage to generate a knockout mutant of the sea urchin homolog of the drug transporter ABCB1, a major player in xenobiotic disposition for all animals. Using CRISPR/Cas9, we generated large fragment deletions of ABCB1 and used these readily detected deletions to rapidly genotype and breed mutant animals to homozygosity in the F2 generation. The knockout larvae are produced according to expected Mendelian distribution, exhibit reduced xenobiotic efflux activity and can be grown to maturity. This study represents a major step towards more sophisticated genetic manipulation of the sea urchin and the establishment of reproducible sea urchin animal resources., Competing Interests: Competing interests The authors declare no competing or financial interests., (© 2022. Published by The Company of Biologists Ltd.)

Rht12, a dominant dwarfing gene of wheat, was shown to be located distally on the long arm of chromosome SA. Lack of recombination with the awn inhibitor BI suggested that Rhtl2 is either tightly linked to this gene or is, in this material, a pleiotropic expression of the gene. Linkage to fi-Amy-A1 was also very tight, indicating that Rhtl2 is present on the segment of chromosome 5AL ancestrally translocated from 4AL. The close linkage to β-Amy-Al also suggests that Rhtl2 is not a homoeoallele of the commercially important GA-insensitive dwarfing genes. Analysis of near-isogenic lines in a number of genetic backgrounds showed that Rhtl2 reduces height without altering ear size and significantly increases spikelet fertility. However its successful utilization in breeding programmes will require careful selection since in some backgrounds the gene reduces grain numbers and grain size. In all backgrounds, Rhtl2 delayed ear emergence time by around 6 days. A delay of this magnitude could, in many environments, adversely affect yield if it is not neutralized by altering the balance of other genes determining ear emergence time. [ABSTRACT FROM AUTHOR]

Vitellogenin (Vg) is important for insect egg maturation and embryo development. In the present study, we characterized the molecular structure and expression profile of Vg gene, and analyzed its reproductive functions in diamondback moth, Plutella xylostella (L.), a destructive pest of cruciferous crops, using CRISPR/Cas9 system. The P. xylostella Vg (Px Vg) included all conserved domains and motifs that were commonly found in most insect Vgs except for the polyserine tract. PxVg gene was highly expressed in female pupae and adults. Px Vg protein was detected in eggs and female adults. PxVg was mainly expressed in the fat body and its protein was detected in most tissues, except in the midgut. CRISPR/Cas9-induced PxVg knockout successfully constructed a homozygous mutant strain with a 5-base pair nucleotide deletion. No Px Vg protein was found in the mutant individuals and in their ovaries. There were no significant differences between wild (WT) and mutant (Mut-5) types of P. xylostella in terms of ovariole length and the number of fully developed oocytes in newly emerged females. No significant difference was observed in the number of eggs laid within two days, but there was a lower egg hatchability (84% for WT vs. 47% for Mut-5). This is the first study presenting the functions of Vg in ovary development, egg maturation, oviposition and embryonic development of P. xylostella. Our results suggest that the reproductive functions of Vg may be species-specific in insects. It is possible that Vg may not be the major egg yolk protein precursor in P. xylostella. Other "functional Vgs" closely involved in the yolk formation and oogenesis would need to be further explored in P. xylostella. Image 1 • PxVg is mainly expressed in female adults, specifically expressed in the fat body. • A homozygous mutant of P. xylostella with 5-bp-nucleotide deletion in the PxVg was developed using CRISPR/Cas9 system. • Knockout of PxVg showed completely inhibition of Vg deposition in the ovaries of mutant individuals. • No significant difference was observed in ovarian development after knockout of PxVg. • No significant difference was observed in oviposition , but there was a lower egg hatchability after knockout of PxVg. [ABSTRACT FROM AUTHOR]

Abstract The zero erucic acid Ethiopian mustard lines developed so far are characterized by an exceptionally high linolenic acid content in the seed oil. The mutant line N2-4961, expressing low linolenic acid content in a high erucic acid background, was developed through chemical mutagenesis. The objective of this research was to study the inheritance of low linolenic acid content in this mutant. Line N2-4961 was reciprocally crossed with its parent line C-101 and the linolenic acid content of the reciprocal F1 , F2 and BC1 generations was studied. No maternal, cytoplasmic or dominance effects were detected in the analysis of F1 seeds and F1 plants from reciprocal crosses. Linolenic acid content segregated in 1 : 2 : 1 ratios in all the F2 populations studied, suggesting monogenic inheritance. This was confirmed with the analysis of the reciprocal backcross generation. The simple inheritance of low linolenic acid content in N2-4961 will facilitate the transference of this trait to zero erucic acid lines of Ethiopian mustard. [ABSTRACT FROM AUTHOR]