Assessment of different anther culture approaches to produce doubled haploids in cucumber (Cucumis sativus L.)

[EN] Cucumber is one of the most important vegetable crops worldwide, which makes it a good candidate to produce doubled haploid (DH) lines to accelerate plant breeding. Traditionally, these approaches involved induction of gynogenesis or parthenogenesis with irradiated pollen, which carries some disadvantages compared to androgenesis. Despite this, studies on anther/microspore cultures in cucumber are surprisingly scarce. Furthermore, most of them failed to unambiguously demonstrate the haploid origin of the individuals obtained. In this work we focused on anther cultures using two cucumber genotypes, different previously published protocols for anther culture, different in vitro culture variants to make it more efficient, and most importantly, a combination of flow cytometry and microsatellite molecular markers to evaluate the real androgenic potential and the impact of anther wall tissue proliferation. We developed a method to produce DH plants involving a bud pretreatment at 4 C, a 35 C treatment to anthers, culture with BAP and 2,4-D, and induction of callus morphogenesis by an additional 35 C treatment and sequential culture first in liquid medium in darkness and second in solid medium with light. We also found that factors such as genotype, proliferation of anther wall tissues, orientation of anthers in the culture medium and growth regulator composition of the initial anther culture medium have a remarkable impact. Our rate of chromosome doubling (81%) was high enough to exclude additional chromosome doubling steps. Together, our results present androgenesis as an improvable but yet more convenient alternative to traditional gynogenesis and parthenogenesis-based approaches.Thanks are due to all the whole staff of the Cell Biology Group for helping and training AA during his stay in the group. This work was supported by Grant AGL2017- 88135-R to JMSS from Spanish Ministerio de Economı´a y Competitividad (MINECO) jointly funded by FEDER.Asadi, A.; Zebarjadi, A.; Abdollahi, MR.; Seguí-Simarro, JM. (2018). Assessment of different anther culture approaches to produce doubled haploids in cucumber (Cucumis sativus L.). Euphytica. 214(216):1-17. https://doi.org/10.1007/s10681-018-2297-xS117214216Abdollahi MR, Najafi S, Sarikhani H, Moosavi SS (2016) Induction and development of anther-derived gametic embryos in cucumber (Cucumis sativus L.) by optimizing the macronutrient and agar concentrations in culture medium. Turk J Biol 40(3):571–579Ashok Kumar HG, Murthy HN (2004) Effect of sugars and amino acids on androgenesis of Cucumis sativus. Plant Cell, Tissue Organ Cult 78(3):201–208. https://doi.org/10.1023/b:ticu.0000025637.56693.68Bai B, Su YH, Yuan J, Zhang XS (2013) Induction of somatic embryos in arabidopsis requires local YUCCA expression mediated by the down-regulation of ethylene biosynthesis. Mol Plant 6(4):1247–1260. https://doi.org/10.1093/mp/sss154Claveria E, Garcia-Mas J, Dolcet-Sanjuan R (2005) Optimization of cucumber doubled haploid line production using in vitro rescue of in vivo induced parthenogenic embryos. J Am Soc Hortic Sci 130(4):555–560Corral-Martínez P, Nuez F, Seguí-Simarro JM (2011) Genetic, quantitative and microscopic evidence for fusion of haploid nuclei and growth of somatic calli in cultured ms1035 tomato anthers. Euphytica 178(2):215–228. https://doi.org/10.1007/s10681-010-0303-zDanin-Poleg Y, Reis N, Tzuri G, Katzir N (2001) Development and characterization of microsatellite markers in Cucumis. Theor Appl Genet 102(1):61–72. https://doi.org/10.1007/s001220051618Dong Y-Q, Zhao W-X, Li X-H, Liu X-C, Gao N-N, Huang J-H, Wang W-Y, Xu X-L, Tang Z-H (2016) Androgenesis, gynogenesis, and parthenogenesis haploids in cucurbit species. Plant Cell Rep. https://doi.org/10.1007/s00299-016-2018-7FAOSTAT (2018) http://faostat.fao.org. Accessed July 2018Ficcadenti N, Sestili S, Annibali S, Di Marco M, Schiavi M (1999) In vitro gynogenesis to induce haploid plants in melon Cucumis melo L. Genet Breed 53:255–257Gałązka J, Niemirowicz-Szczytt K (2013) Review of research on haploid production in cucumber and other cucurbits. Folia Hortic. https://doi.org/10.2478/fhort-2013-0008Hamidvand Y, Abdollahi MR, Chaichi M, Moosavi SS (2013) The effect of plant growth regulators on callogenesis and gametic embryogenesis from anther culture of cucumber (Cucumis sativus L.). Int J Agric Crop Sci 5(10):1089Kurtar ES, Balkaya A, Kandemir D (2016) Evaluation of haploidization efficiency in winter squash (Cucurbita maxima Duch.) and pumpkin (Cucurbita moschata Duch.) through anther culture. Plant Cell, Tissue Organ Cult 127(2):497–511. https://doi.org/10.1007/s11240-016-1074-6Lotfi M, Alan AR, Henning MJ, Jahn MM, Earle ED (2003) Production of haploid and doubled haploid plants of melon (Cucumis melo L.) for use in breeding for multiple virus resistance. Plant Cell Rep 21(11):1121–1128Metwally EI, Moustafa SA, El-Sawy BI, Shalaby TA (1998) Haploid plantlets derived by anther culture of Cucurbita pepo. Plant Cell, Tissue Organ Cult 52(3):171–176. https://doi.org/10.1023/a:1005908326663Mohamed M, Refaei E (2004) Enhanced haploids regeneration in anther culture of summer squash (Curcurbita pepo L.). Cucurbit Genet Coop Rep 27:57–60Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–479Parra-Vega V, Renau-Morata B, Sifres A, Seguí-Simarro JM (2013) Stress treatments and in vitro culture conditions influence microspore embryogenesis and growth of callus from anther walls of sweet pepper (Capsicum annuum L.). Plant Cell, Tissue Organ Cult 112(3):353–360. https://doi.org/10.1007/s11240-012-0242-6Rakha M, Metwally E, Moustafa S, Etman A, Dewir Y (2012) Evaluation of regenerated strains from six Cucurbita interspecific hybrids obtained through anther and ovule in vitro cultures. Aust J Crop Sci 6(1):23–30Saghai-Maroof MA, Soliman KM, Jorgensen RA, Allard RW (1984) Ribosomal DNA spacer-length polymorphisms in barley: mendelian inheritance, chromosomal location, and population dynamics. Proc Natl Acad Sci USA 81(24):8014–8018. https://doi.org/10.1073/pnas.81.24.8014Sauton A, Dumas de Vaulx R (1987) Obtention de plantes haploides chez melon (Cucumis melo L.) par gynogenese indute par du pollen irraidié. Agronomie 7:141–148Seguí-Simarro JM (2016) Androgenesis in solanaceae. In: Germanà MA, Lambardi M (eds) In vitro embryogenesis. Methods in molecular biology, vol 1359. Springer, New York, pp 209–244. https://doi.org/10.1007/978-1-4939-3061-6_9Seguí-Simarro JM, Nuez F (2006) Androgenesis induction from tomato anther cultures: callus characterization. Acta Hort 725:855–861Seguí-Simarro JM, Nuez F (2007) Embryogenesis induction, callogenesis, and plant regeneration by in vitro culture of tomato isolated microspores and whole anthers. J Exp Bot 58(5):1119–1132Seguí-Simarro JM, Nuez F (2008) Pathways to doubled haploidy: chromosome doubling during androgenesis. Cytogenet Genome Res 120(3–4):358–369. https://doi.org/10.1159/000121085Shalaby TA (2006) Embryogenesis and plantlets regeneration from anther culture of squash plants (Cucurbita pepo L.) as affected by different genotypes. J Agric Res Tanta Univ 32(1):173–183Song H, Lou QF, Luo XD, Wolukau JN, Diao WP, Qian CT, Chen JF (2007) Regeneration of doubled haploid plants by androgenesis of cucumber (Cucumis sativus L.). Plant Cell, Tissue Organ Cult 90(3):245–254. https://doi.org/10.1007/s11240-007-9263-ySteward FC, Mapes MO, Mears K (1958) Growth and organized development of cultured cells. II. Organization in cultures grown from freely suspended cells. Am J Bot 45(10):705–708Su YH, Zhao XY, Liu YB, Zhang CL, O’Neill SD, Zhang XS (2009) Auxin-induced WUS expression is essential for embryonic stem cell renewal during somatic embryogenesis in arabidopsis. Plant J 59(3):448–460. https://doi.org/10.1111/j.1365-313X.2009.03880.xSuprunova T, Shmykova N (2008) In vitro induction of haploid plants in unpollinated ovules, anther and microspore culture of Cucumis sativus. In: Cucurbitaceae 2008: proceedings of the IXth Eucarpia meeting on genetics and breeding of cucurbitaceae, pp 371–374Xie M, Qin L-Y, Pan J-S, He H-L, Wu A-Z, Cai R (2005) Flower morphogenesis and microspore development versus anther culture of cucumber. Acta Bot Boreal-Occid Sin 25(6):1096Zhan Y, Chen J-F, Malik AA (2009) Embryoid induction and plant regeneration of cucumber (Cucumis sativus L.) through microspore culture. Acta Hort Sin 36(2):221–22

Allelopathic Effects of Aqueous Extract of Leaf Stem and Root of Sorghum bicolor on Seed Germination and Seedling Growth of Vigna radiata L.

Seed germination under field conditions is highly influenced by the presence of other plants. Allelopathy is an important mechanism of plant competition, by producing phytotoxins to the plant environment in order to decline other plants� growth. Soil sickness problem in farm lands is also known as an allelopathic effect or even autotoxicity. The toxicity of released allelochemicals by a plant in the environment is attributed to its function of concentration, age and metabolic stage. In this study we investigate the effect (5, 20, 35 and 50 g l-1) of leaf, stem and root water extract of sorghum on seed germination and seedling growth of mung bean. The results of the experiment showed that allelopathic effect of different concentrations was not significant for germination percentage, but germination rate and mean germination time decreased significantly by increasing the concentration of allelopathic extracts; also, there was a clear allelopathic effect of sorghum extract on seedling growth of mung bean. 50 g l-1 sorghum stem extract exhibited the highest inhibitory effect on root and shoot growth of mung bean. Among all parts of sorghum, stem extracts showed the highest allelopatic effect on seedling growth. Root extract showed higher inhibitory effect than leaf extracts

The relationship of sexual dysfunction disorders syndrome and body image with mental health in women

BACKGROUND: Sexual dysfunction (SD) and body image are interrelated and could affect mental health. This study aims to examine the relationship between SD and body image with mental health in women.METHODS: This was a correlational study and statistical population was all married women at age of 25-50 who referred to gynecology centers in Tehran, Iran, in Mar 2015. 150 women who referred to gynecological clinics in two regions of 4 and 8 in Tehran, were chosen using convenience sampling method. They filled 12-Item General Health Questionnaire (GHQ-12), Sexual Function Questionnaire (SFQ), and Body Image Scale (BIS). Data were analyzed by using Pearson correlation and regression through SPSS software.RESULTS: The relationship between SD disorders syndrome and negative body image with mental health was statistically significant (P < 0.001).CONCLUSION: Negative self-image could have a negative effect on SD syndrome and it could predict poor mental health in women