Effect of Supplementary Irrigation on the Yield of Sorghum (Sorghum bicolor L. Moench) in the Context of Climate Change in the Dry Savannahs of Togo

Under the current context of climate change, supplementary irrigation may be needed for crop production resilience. We determined the effects of supplementary irrigation on sorghum grain yield in the dry Savannah region of Togo. A two-year trial was conducted in a controlled environment at AREJ, an agro-ecological center in Cinkassé. The plant material was sorghum variety Sorvato 28. The experimental design was a Completely Randomized Block with three replications and three treatments as follows: T0 control plot (rainfed conditions);T1 (supplementary irrigation from flowering to grain filling stage) and T2 (supplementary irrigation from planting to grain filling stage). Two irrigation techniques (furrow and Californian system) were used under each watering treatment. The results showed that irrigation technique significantly affected panicle length with no effect on 1000 grains mass. Panicle length and grain yields varied from 15.59 to 25.71 cm and 0.0 to 2.06 t∙h−1, respectively, with the highest values (25.66 cm and 2.06 t∙h−1, respectively) under the T2 treatment with the California system-based supplementary irrigation. The comparison of results obtained on treatment T0 and T2, shows that supplementary irrigation increased the yields by at least 68.62%. Supplementary irrigation during sowing and growing season (T2) improved sorghum yields in the dry savannahs of Togo, with a better performance of the California irrigation system.

高粱(Sorghum bicolor(L.)Moench)种质资源研究进展

高粱(Sorghum bicolor(L.)Moench)是世界上主要的旱粮作物,也是全球5亿多人口的主要粮食。文章对高粱的起源与分类、高粱种质资源收集、保存和利用以及遗传多样性研究与核心种质的构建等方面的研究进行了综述,最后对我国高粱种质资源的研究方向进行了展望。

旱稻(Oryza sativa)/稗草(Echinochloa caudata)//高梁(Sorghum bicolor)三属杂种表现及分析

以旱稻基因型远F2 1 [旱 65 (Oryzasativa) 长芒稗 (Echinochloacaudata) ]为母本 ,高粱(Sorghumbicolor)基因型沈农 1 33为父本 ,进行属间远缘杂交 ,获得三属间杂交实粒种子。对杂交种F0 、F1、F2 代连续观察结果表明 :( 1 )杂交F0 代高度不孕 ,并伴有杂交种发育夭折现象发生 ,结实率仅为 2 63‰ ;( 2 )F1杂种优势明显 ,株高、穗长、1级枝梗数、小花数、结实粒数、结实率、千粒重诸性状超母本优势为 0 61 %～ 2 0 5 6% ;主茎叶片数多于母本 1片 ,总叶面积超亲优势为 1 1 95 % ;F1出现母本没有的红芒、红护颖和紫柱头 3个可遗传性状 ;( 3)F2 除在生育期、株高及穗长等性状发生分离外 ,其典型特征是穗型、芒性、芒色、柱头颜色。

建立甜高粱(Sorghum bicolor)高频、高效再生体系的研究

【目的】建立能源植物甜高粱(Sorghum bicolor L.Moench)的高频、高效离体培养再生体系,为遗传转化奠定基础。【方法】以甜高粱品种凯勒的芽顶端分生组织为外植体,探讨在甜高粱直接诱导丛生芽的培养过程中,无菌实生苗的适合苗龄,适宜的激素组合与比例;并将获得的最佳培养方法应用于其它两个甜高粱品种:M-81E和意达利,以研究此途径的基因型依赖性。【结果】利用3d苗龄的芽顶端分生组织作为外植体容易获得较高的分生组织膨大率。甜高粱丛生芽诱导的适宜激素组合为4.0mg·L-16-苄基腺嘌呤+0.5mg·L-12,4-二氯苯氧乙酸+0.5mg·L-1噻重氮苯基脲,可获得91%的丛生芽诱导频率。适合甜高粱快速生根的激素配方为0.5mg·L-1萘乙酸。通过芽顶端分生组织直接诱导丛生芽途径,每个外植体可诱导出上百个芽,最终产生30—40株再生苗。M-81E和意达利具有与凯勒相似的丛生芽诱导频率及效率。【结论】本研究成功地建立了甜高粱的高频、高效离体培养再生体系,且具有外植体来源不受限制,基因型依赖性弱,遗传稳定性好的优点。

高粱[Sorghum bicolor(L.)Moench.]叶面积校正值(K 值)的初步研究

通过对两个高粱品种叶面积校正值(K 值)的初步研究,提出了在高粱叶面积测算中采用叶面积校正值K=0.72的建议,并对K 值在应用中的可靠性进行了分析。

高粱(Sorghum bicolor)分子图谱的构建及寄生草(Striga asiatica)萌发诱导物基因的定位

高粱是世界上第五大主要粮食作物，也是非洲国家的主要粮食来源之一，Ｓｔｒｉｇａ ａｓｉａｔｉｃａ是一种寄生于高粱等主要农作物的野生草之一。选用两个对寄生草抗性表现差异的 高粱品系“山桂红”和“ＳＲＮ３９”作亲本，构建了一个重组近交系群体（Ｒｅｃｏｍｂｉｎａｎｔ ｉｎｂｒｅｄ， ＲＩ），并随机筛选出９４个系用于构建分子连锁图谱和基因定位。在应用的２８６个多态性标记 中，有２５１个标记分别标定在１０条不同的连锁群上，标记间的平均图距为７．１ｃｍ，总图谱覆盖 了高粱基因组的１７７９ｃｍ，是目前国际上几个比较完整的高粱分子连锁图谱之一。群体的共 分离分析表明，与寄生草抗性有关的萌发诱导物基因（Ｇｅｒｍｉｎｉｎａｔｉｏｎ ｓｔｉｍｕｌａｎｔ ｇｅｎｅ－ＧｅｒｍＳｔｉｍ） 位于高粱的遗传连锁群Ｊ上，相距较近的分子标记为１３ｃｍ。进一步的精细定位分析，发现有 两个分子标记分别位于基因的两侧，距离为１．６和２．１个ｃｍ。

Effects of Planting Density and Row Spacing on Grass Yield of Forage Sweet Sorghum(Sorghum bicolor [L.]Moench)

[Objective] The aim was to explore high-yielding cultivation techniques for forage sweet sorghum. [Method[ The effects of planting density and row spacing on plant productivity and grass yield of forage sweet sorghum （Sorghum bicolor （L.） Moench） were compared using split-plot design and LSD method of IBMSPSSStatis- ticsv22. [Result]The planting density and row spacing had important influence on the plant productivity and yield of forage sweet sorghum. The optimum planting density- row spacing combination for plant productivity of forage sweet sorghum was A1B,, i. e., planting density of 75 000 plants/hm2 and row spacing of 40 cm, and the opti- mum combination for yield of forage sweet sorghum was A2B,, i.e., planting density of 225 000 plants/hm2 and row spacing of 40 cm. [Conclusion] This study will pro- vide theoretical basis and technical support for the production practice of forage sweet sorghum.

QTLs for Sugar Content of Stalk in Sweet Sorghum(Sorghum bicolor L.Moench)

High sugar content of sorghum stalk is an important factor in the sorghum silage production. To identify the genomic regions controlling sugar content and to develop molecular markers linked to sugar content in sweet sorghum, we used an F2：3 segregating population consisting of 207 individuals derived from a cross between a high sugar content inbred line, Early Folger, and a normal inbred line, N32B, for genetic linkage mapping and quantitative trait locus （QTL） analysis. We constructed a genetic linkage map spanning 983.5 cM based on a total of 327 markers comprising 31 restriction fragment length polymorphism （RFLP） markers, 254 amplified fragment length polymorphism （AFLP） markers, and 42 simple sequence repeat （SSR） markers. In the 20 linkage groups detected, 98.2% of markers aligned to the 10 linkage groups of sorghum. Variations in sugar content at different growth stages and among internodes suggested that the sugar content of middle internodes is stable and suitable for measuring at early dough stage. The broad sense heritability （hB0 of sugar content was 0.64 and 0.62 estimated from the data of F3 families and each parent in 2003 and 2004. We identified one and two QTLs accounting for 22.2 to 25.0% of phenotypic variance using simple interval mapping method in 2003 and 2004, respectively. These two QTLs showed a negative additive effect, and over-dominance effect. A QTL on LG-D was detected in both two years. Above results will be help us to understand the genetic mechanism of sugar content in sorghum and the QTL detected in this study might be useful in the improvement of sugar content by marker-assisted selection.

Genome-wide association study for starch content and constitution in sorghum(Sorghum bicolor(L.) Moench)

Starch is the most important component in endosperm of sorghum grain.Usually,two types of starch are present:amylose(AM)and amylopectin(AP).The levels of AM and AP contents play a significant role in the appearance,structure,and quality of sorghum grains and in marketing applications.In the present study,a panel of 634 sorghum(Sorghum bicolor(L.)Moench)accessions were evaluated for starch,AM,and AP contents of grain,which included a mini core collection of 242 accessions from the International Crops Research Institute for the Semi-Arid Tropics(ICRISAT)in India,and 252 landraces and 140 cultivars from China.The average starch content was 67.64%and the average AM and AP contents were 20.19 and 79.81%,respectively.We developed a total of 260000 high-confidence single nucleotide polymorphism(SNP)markers in the panel of 634 accessions of S.bicolor using specific locus amplified fragment sequencing(SLAF-seq).We performed genome-wide association studies(GWAS)of starch,AM,and AM/AP of grain and SNP markers based on a mixed linear model(MLM).In total,70 significant association signals were detected for starch,AM,and AM/AP ratio of grain with P<4.452×10^-7,of which 10 SNPs were identified with significant starch,51 SNPs were associated with AM,and nine SNPs were associated with the AM/AP ratio.The Gene Ontology(GO)analysis identified 12 candidate genes at five QTLs associated with starch metabolism within the 200-kb intervals,located on chromosomes 1,5,6,and 9.Of these genes,Sobic.006G036500.1 encodes peptidyl-prolyl cis-trans-isomerase CYP38 responsible for hexose monophosphate shunt(HMS)and Sobic.009G071800 encodes 6-phospho-fructokinase(PFK),which is involved in the embden-meyerhof pathway(EMP).Kompetitive allele specific PCR(KASP)markers were developed to validate the GWAS results.The C allele is correlated with a high starch content,while the T allele is linked with a low level of starch content,and provides reliable haplotypes for MAS in sorghum quality improvement.

高粱(Sorghumbicolor(L.)Moench)未成熟胚乳培养的研究

高粱未成熟胚乳在含有 BA或 2 .4-D与 KT组合的培养基上诱导启动 .诱导形成愈伤组织的适宜激素是 :2 .4-D4mg/L,BA1m g/L,NAA0 .2 mg/L 的组合或 2 .4-D2 mg/L,KT1m g/L 的组合 .适宜的分化激素组合是 (1)2 .4-D0 .5 mg/L,NAA0 .2 mg/L,BA0 .7mg/L,KT4mg/L;(2 ) NAA0 .5 mg/L,ZT1mg/L;(3 ) 2 .4-D0 .5 m g/L,KT1m g/L.脱分化后的胚乳细胞 ,部分细胞进入分裂期形成愈伤组织 ,胚乳愈伤组织有较强的分化能力 .植株形成方式为器官发生 .

Vacuolar invertase genes SbVIN1 and SbVIN2 are differently associated with stem and grain traits in sorghum (Sorghum bicolor)

In higher plants, vacuolar invertases play essential roles in sugar metabolism, organ development, and sink strength. In sorghum(Sorghum bicolor), two vacuolar invertase genes,Sb VIN1(Sobic. 004 G004800) and Sb VIN2(Sobic. 006 G160700) have been reported, but their enzymatic properties and functional differences are largely unknown. We combined molecular, biochemical and genomic approaches to investigate their roles in sorghum stem and grain traits. Sb VIN1 and Sb VIN2 showed different expression levels in internodes,leaves, and panicles, indicating that their importance in each organ was different. In an in vitro sucrose hydrolysis assay, proteins of both genes heterologously expressed in Pichia pastoris displayed similar enzyme properties including the same optimum reaction p H(5)and similar Kmfor sucroe(49 mmol L-1 and 45 mmol L-1 for Sb VIN1 and Sb VIN2,respectively). The optimum reaction temperatures of Sb VIN1 and Sb VIN2 were 45 °C and65 °C, respectively. Sb VIN2 showed higher tolerance to high temperature than Sb VIN1. We characterized the sequence variation of these two vacuolar invertase genes in a panel of 216 diverse inbred lines of sweet and grain sorghum and performed gene-based association analysis. Sb VIN1 showed significant associations with stem traits including stem length,stem diameter, internode number, stem fresh weight, and Brix, as well as grain traits including hundred-grain weight and grain width. Significantly associated variation sites were mainly in 5′ upstream and intron regions. Sb VIN2 only associated with grain width and stem water-soluble carbohydrates(WSCs) content. We conclude that the vacuolar invertase genes Sb VIN1 and Sb VIN2 are differently associated with stem and grain traits in sorghum.

Evaluation of Plant Densities and Various Irrigation Regimes of Sorghum (Sorghum bicolor L.) under Low Water Supply

Drought stress, during growth season along with plant density, is an important problem that needs attention. In order to investigate the influence of both factors in increasing the water use efficiency, field experiments were laid out in split-plot design at Agriculture Research Station, Collage of Food and Agriculture Sciences, King Saud University, to investigate the effects of irrigation intervals viz., irrigation every (6, 9 and 12 days) under different plant densities i.e., (6, 8 and 10 plants/m2) on growth, yield and yield component parameters as well as grain quality of sorghum local variety (Gizani). Results revealed that almost all growth, yield and yield component parameters were significantly influenced by both factors as well as their interaction. Chemical composition of seeds, leaf proline content and WUE were also considered. Severe drought stress condition caused gradual decrease in most of the growth characters as compared to watered treatment and reflected in decreasing yield and yield component characters. Increasing plant densities led to raise biomass production and seed yield per unit area and not able to compensated the low number and weight of grains per panicle. Contrary, low plant density, under adequate irrigation conditions, can be compensated by a high number of grains per panicle and high weight of the grain. Maximum seed yield per hectare was recorded by the interactional effects of most watered treatments (irrigation every 6 days) and plant density of 10 plants per square meter.

Genetic Variation among Sorghum (<i>Sorghum bicolor</i>L. Moench) Landraces from Eritrea under Post-Flowering Drought Stress Conditions

Sorghum (Sorghum bicolour (L.) Moench) grown under rain-fed conditions is usually affected by drought stress at different stages, resulting in reduced yield. The assessment of variation in morpho-physiological traits contributing towards drought tolerance at these stages is of vital importance. This study was conducted using a split plot design with three replications to evaluate 25 sorghum accessions at post flowering stage under well watered and drought stress conditions at Hamelmalo Agricultural College. The data of 14 different morpho-physiological traits were subjected to analysis of variance, estimation of genetic variability and heritability and principal component analysis. We analyzed variance for seedling vigor, number of leaves, leaf area, stay-green, peduncle exsertion, panicle length and width, plant height, days to flowering and maturity, grain yield, biomass and harvest index under drought stress and irrigated conditions. The results showed that genotypic differences were significant at P 1 explaining 74.6% of the total variation with grain yield, biomass, stay-green, leaf area, peduncle exsertion and days to flowering and maturity being the most important characters in PC1 and PC2. This research demonstrated high diversity for the characters studied. Moreover, the result showed that drought stress reduced the yield of some genotypes, though others were tolerant to drought. Accessions EG 885, EG 469, EG 481, EG 849, Hamelmalo, EG 836 and EG 711 were identified as superior for post-flowering drought tolerance and could be used by breeders in improvement programs.

Differences in Nuclear DNA Between Male-Sterile and Male-Fertile Lines of Sorghum bicolor

Cytoplasmic male sterility(cms)is determined by nuclear-cytoplasmic interactions. Up to now, most studies are focused on the comparison of cytoplasmic DNAs of male-sterile lines and male-fertile lines, and analysis of nuclear DNA has not been documented yet. In order to find out the possible difference in nuclear genome of male-sterile line A1 Tx623 and corresponding male-fertile line Tx623 of sorghum, random amplified polymorphic DNA(RAPD)approach was used to analyze their cytoplasmic and nuclear genomes. Total DNAs of them were amplified at first to screen primers, which were able to generate reproducible bands specific to male-sterile line or male-fertile line. Then the selected primers were used to amplify their mitochon-drial DNA(mtDNA)and chloroplast DNA(cpDNA). The origins of all the polymorphic fragments were analyzed. After ruling out those amplified from cytoplasmic DNA, seventeen polymorphic fragments were determined to be amplified from nuclear DNA. These fragments originated from nuclear DNA indicate that differences in sequence exist between the nuclear DNA of male-sterile line and male-fertile line of sorghum, which do not agree with the traditional standpoint that they have identical nucleus.

Characterization of subunits encoded by Sn RK1 and dissection of combinations among these subunits in sorghum(Sorghum bicolor L.)

Sucrose nonfermenting-related protein kinase 1(SnRK1)is one of the critical serine/threonine protein kinases.It commonly mediates plant growth and development,cross-talks with metabolism processes and physiological responses to biotic or abiotic stresses.It plays a key role in distributing carbohydrates and sugar signal transporting.In the present study,eight SnRK1 coding genes were identified in sorghum(Sorghum bicolor L.)via sequences alignment,with three forαsubunits(SnRK1α1 to SnRK1α3),three forβ(SnRK1β1 to SnRK1β3),and one for bothγ(SnRK1γ)andβγ(SnRK1βγ).These eight corresponding genes located on five chromosomes(Chr)of Chr1–3,Chr7,and Chr9 and presented collinearities to SnRK1s from maize and rice,exhibiting highly conserved domains within the same subunits from the three kinds of cereals.Expression results via qRT-PCR showed that different coding genes of SnRK1s in sorghum possessed similar expression patterns except for SnRK1α3 with a low expression level in grains and SnRK1β2 with a relatively high expression level in inflorescences.Results of subcellular localization in sorghum leaf protoplast showed that SnRK1α1/α2/α3/γmainly located on organelles,while the rest four of SnRK1β1/β2/β3/βγlocated on both membranes and some organelles.Besides,three combinations were discovered among eight SnRK1 subunits in sorghum through yeast two hybrid,includingα1-β2-βγ,α2-β3-γ,andα3-β3-γ.These results provide informative references for the following functional dissection of SnRK1 subunits in sorghum.

Juice, Ethanol, and Grain Yield Potential of Five Sweet Sorghum (<i>Sorghum bicolor</i>[L.] Moench) Cultivars

Sweet sorghum (Sorghum bicolor [L.] Moench) accumulates fermentable sugars in the stem and is increasingly being studied as a potential source of feedstock for bioethanol production. The objective of this study was to evaluate biomass and grain yield in five sweet sorghum cultivars (Dale, M81E, Sugar Drip, Della and Keller) and to determine quality of extractable juice and grain. Randomized complete block experiments were performed in the summer of 2009, 2010, and 2011. Leaf dry weight varied with year and cultivar and averaged 6177 kg·ha-1. Fresh stem weight ranged from 21 to 54 Mg·ha-1 with a mean across years and cultivars of 32.9 Mg·ha-1. Variations in stem weight were correlated with extractable juice volumes that ranged from 10 to 24 m3·ha-1. Juice Brix values fell within a narrow range (14% - 19%) across years and cultivars with an average of 15.6%. In all production years, theoretical sugar and ethanol yield were always numerically higher for Keller and M81E. Grain yield was lowest in Keller (90 kg·ha-1), but ranged from 400 to 1300 kg·ha-1 in other cultivars with a mean of 584 kg·ha-1 across years. However, Keller had the highest starch content with a lower proportion of resistant starch in the grain. Except for Keller, the cultivars tested are potential sources of both fermentable sugars and grain.

Effect of Sowing Rate on Agronomic Traits of Sorghum (Sorghum bicolor Moench) in Southern Kyushu, Japan

Newly released sudangrass-type sorghum cv. “Tarzan” was bred for use as a bioenergy-resource feedstuff in Germany. Since the genotype was collected at a high altitudinal site in the Central Africa continent, its early growth is vigorous at low temperature. As vigorous growth at low temperature is derived from high tillering ability, the sowing rate of this genotype should be reduced from the ordinary rate for sorghum. Thus, in the present study, the optimal sowing rate of cv. “Tarzan” in southern Kyushu was determined by the effect of sowing rate at 0.5, 1.0, and 1.5 g·m-2, designated as low (L), middle (M), and high (H) levels, respectively, on growth habit compared with the normal sudangrass cultivar “Roll-king II”. Early growth was retarded by heavy precipitation in June, after which the growth was comparable for the two genotypes. Dry matter yield of “Tarzan” increased with higher sowing rate;the H level yield was 1298 g·m-2, and the yield was not adversely affected by the humidity and heat in the summer growing season, compared with “Roll-king II”. This suggests that “Tarzan” can adapt to the humid, hot climate in southern Kyushu, and the optimum sowing rate of “Tarzan” should be as low as 1.0 - 1.5 g·m-2, which is less than one-fifth to one-third that of forage sudangrass grown in the region.

Genome-wide identification and characterization of the JAZ gene family and its expression patterns under various abiotic stresses in Sorghum bicolor

The jasmonate ZIM domain(JAZ)protein belongs to the TIFY((TIF[F/Y]XG)domain protein)family,which is composed of several plant-specific proteins that play important roles in plant growth,development,and defense responses.However,the mechanism of the sorghum JAZ family in response to abiotic stress remains unclear.In the present study,a total of 17 JAZ genes were identified in sorghum using a Hidden Markov Model search.In addition,real-time quantification polymerase chain reaction(RT-qPCR)was used to analyze the gene expression patterns under abiotic stress.Based on phylogenetic tree analysis,the sorghum JAZ proteins were mainly divided into nine subfamilies.A promoter analysis revealed that the SbJAZ family contains diverse types of promoter cis-acting elements,indicating that JAZ proteins function in multiple pathways upon stress stimulation in plants.According to RT-qPCR,SbJAZ gene expression is tissuespecific.Additionally,under cold,hot,polyethylene glycol,jasmonic acid,abscisic acid,and gibberellin treatments,the expression patterns of SbJAZ genes were distinctly different,indicating that the expression of SbJAZ genes may be coordinated with different stresses.Furthermore,the overexpression of SbJAZ1 in Escherichia coli was found to promote the growth of recombinant cells under abiotic stresses,such as PEG 6000,NaCl,and 40℃ treatments.Altogether,our findings help us to better understand the potential molecular mechanisms of the SbJAZ family in sorghum in response to abiotic stresses.

Improved Culture Media for Embryogenic Callus Generation in Sorghum [Sorghum bicolor (L.) Moench]

Many attempts on optimization of sorghum[Sorghum bicolor(L.)Moench]tissue culture induction media have been made,but the culture system remains with some bottlenecks compared to that of other crops.This study aimed at assessing the suitability of various induction media to produce embryogenic callus(yellow and friable)with high induction rates and reduced phenolic exudation.The six culture medium modifications:3 based on Murashige and Skoog(MS)medium and one each based on Chu N6,Gamborg B5 and 190-2 media respectively were applied in the culture of mature embryos from 10 sorghum genotypes.Although there was a genotype influence on the attainment of a yellow callus,friability of the callus was determined to be dependent on the culture medium and not the genotype.Half strength MS medium with 0.2 mg/l 2,4-D with 2.8 g/l Gelrite®as the gelling agent modified with 1.0 g/l KH_(2)PO_(4),1.0 g/l L-proline,1.0 g/l L-asparagine and 0.16 mg/l CuSO_(4)·5H_(2)O(type E)was found to be the most effective resulting in about 60%yellow coloured callus induction with 25%friability.Addition of CuSO_(4)·5H_(2)O,KH_(2)PO_(4),L-proline and L-asparagine significantly reduced the phenolic production.Half strength MS medium was observed to contribute to quality callus production when compared to full strength MS media modified with the compounds.The half strength MS medium was also observed to suppress phenolic production.Medium 190-2 produced the highest regeneration frequency(40%)among the 3-regeneration media tested.The results provide information on a suitable sorghum callus induction medium necessary for embryogenesis.

Doubled Haploid Production Using an Improved Anther Culture Protocol for Sorghum[Sorghum bicolor(L.)Moench]

Sorghum[Sorghum bicolor(L.)Moench]can benefit from accelerated breeding and release of improved varieties through doubled haploid technology.The technology has been used in speeding up the breeding of other major cereals such as wheat,maize and rice,for which generally widely applied optimised protocols exist.A reproducible protocol for the crop,that can overcome genotype dependency and other species-specific challenges such as phenolic exudation is however lacking.This study aimed at sorghum doubled haploids production thereby contributing to the development of an improved protocol.From the 28 hybrid genotypes,both F1 registered-and experimental hybrids involved,this study successfully produced haploids from five genotypes and subsequently,four confirmed doubled-haploid lines on W14mf medium or its modification with 1.0 gl^(-1)L-proline,1.0 gl^(-1)L-asparagine and 1.0 gl^(-1)KH_(2)PO_(4).Medium 190-2Cu was used for regeneration and rooting,which occurred successfully,if the calli were transferred on to it less than 7 days after induction,and temperature was maintained at 25°C under light condition.Genotype dependency was not wholly overcome;however,sorghum’s high tillering ability and abiotic stress tolerance were observed to contribute to attainment of haploid plantlets.Spontaneous diploids producing seeds at rates of upto 80.5%were obtained,therefore eliminating the need for colchicine duplication.