MicroRNA expression profiles in response to drought stress in Sorghum bicolor

The regulatory role of small non-coding RNAs that are 20–24 nucleotides in length has become the foremost area of research for biologists. A major class of small RNAs represented by the microRNAs (miRNAs), has been implicated in various aspects of plant development including leaf pattering, meristem function, root patterning etc. Recent findings support that miRNAs are regulated by drought and other abiotic stresses in various plant species. In this study, were report the expression profiling of 8 known abiotic stress deregulated miRNAs in 11 elite sorghum genotypes, under watered and drought conditions. Significant deregulation was observed with miR396, miR393, miR397-5p, miR166, miR167 and miR168. Among these, the expression levels of sbi-miR396 and sbi-miR398 were the highest in all the genotypes. The expression of sbi-miR396 was maximum in the grain sorghum HSD3226 under well-watered conditions and the profile shifted towards HSD3221 under drought stress. Forage accessions, N98 and Atlas, showed an opposite behavior in expression patterns of miR397-5p in drought physiologies. Such dynamic expression patterns could be indicative of prevailing drought tolerant mechanisms present in these sorghum accessions. This data provides insights into sorghum miRNAs which may have potential use in improving drought tolerance in sorghum and other cereal crops.     

Expression of a boiling-stable protein (BsCyp) in response to heat shock, drought and ABA treatments in Sorghum bicolor

The effect of heat stress, drought and abscisic acid (ABA) on the induction of boiling-stable proteins were studied in 12 h germinated sorghum embryos and endosperm. SDS-PAGE revealed the heat shock induction of one major 97 kDa boiling-stable protein both in embryos and endosperm. Western blotting analysis, using polyclonal antibodies raised against a 20 kDa Arabidopsis thaliana cyclophilin, revealed the induction of a cross-reacting band (97 kDa, BsCyp) in both embryos and endosperm in a time dependent manner. The BsCyp boiling-stable polypeptide was also regulated by exogenous application of ABA, indicating its role in water stress adaptation. Drought stress also resulted in a significant induction of boiling-stable protein (BsCyp) in both embryos and endosperm. Based upon these observations the possible role of BsCyp in water stress tolerance is discussed.     

Expression analysis of a stress-modulated transcript in drought tolerant and susceptible cultivars of sorghum (Sorghum bicolor)

The present study reports the cloning of a 581 bp sequence, designated as SbEST8, from the osmotically stressed germinated seeds of a drought tolerant cultivar of sorghum (Sorghum bicolor). The SbEST8, which shows no homology with the reported gene sequences, is present in multiple copies and lacks restriction fragment length polymorphism among different sorghum cultivars. The expression of SbEST8 in the germinating seeds of sorghum was modulated by different abiotic stresses. Kinetic studies revealed that imposition of osmotic stress after 8h resulted in maximum levels of SbEST8 mRNA in the germinating seeds of cv. ICSV-272, with further stress causing a decline to undetectable levels by 16 h. However, relieving the stress after 12h resulted in an enhancement of SbEST8 mRNA levels for at least another 4h following which it declined. The decrease in SbEST8 mRNA levels in the leaves at 30 DAS in response to drought stress was observed only in the drought susceptible cultivar (CSV-216), whereas its expression was either increased substantially or remained unaffected in the tolerant cultivars, thus suggesting its role in water stress tolerance.     

Genotypic variation in the photosynthetic competence of Sorghum bicolor seedlings subjected to polyethylene glycol-mediated drought stress

Eleven varieties of Sorghum bicolor, subjected to PEG-mediated drought stress were compared for their photosynthetic performance. The varieties differed in their relative water content over a range of PEG concentrations (0-25%). CO2 assimilation, stomatal conductance and the quantum yield of PSII electron transport decreased with increasing PEG concentrations in all varieties. However the intercellular CO2 concentration showed a nonlinear PEG concentration-dependent change. At lower PEG concentrations there was a decrease in the levels of intercellular CO2 concentration in all varieties that could be attributed to stomatal closure. At higher PEG concentrations, some varieties showed an increase in the intercellular CO2 concentration, indicating an inhibition of photosynthetic activity due to non-stomatal effects, while others did not. It was seen that the varieties differed in the stress thresholds at which stomatal and metabolic limitations to photosynthesis occur. These differences in the photosynthetic adaptation of Sorghum varieties could be useful in identifying genotypes showing large differences in photosynthetic adaptation, which could be useful in mapping photosynthetic traits for drought stress tolerance.     

Genetic transformation of Sorghum bicolor

Great millet (Sorghum bicolor (L.) Moench) is cultivated across the world for food and fodder. It is typically grown in semiarid regions that are not suitable for cultivation of other major cereals. Sexual incompatibility and shortage of available genes in germplasm to combat biotic and abiotic stresses resulted in marginalized yields of this crop. Genetic modification of sorghum with agronomically useful genes can address this problem. Here, we tried to review and summarize the key aspects of sorghum transformation work being carried out so far by various research groups across the world. The approaches used and the obstacles in generating transgenic sorghum are also pointed out and discussed.     

Mycorrhizal symbiosis and response of sorghum plants to combined drought and salinity stresses

Arbuscular mycorrhizal (AM) symbiosis can confer increased host resistance to drought stress, although the effect is unpredictable. Since AM symbiosis also frequently increases host resistance to salinity stress, and since drought and salinity stress are often linked in drying soils, we speculated that the AM influence on plant drought response may be partially the result of AM influence on salinity stress. We tested the hypothesis that AM-induced effects on drought responses would be more pronounced when plants of comparable size are exposed to drought in salinized soils. In two greenhouse experiments, several water relations characteristics were measured in sorghum plants colonized by Glomus intraradices (Gi), Gigaspora margarita (Gm) or a mixture of AM species, during a sustained drought following exposure to salinity treatments (NaCl stress, osmotic stress via concentrated macronutrients, or soil leaching). The presence of excess salt in soils widened the difference in drought responses between AM and nonAM plants in just two instances. Days required for plants to reach stomatal closure were similar for Gi and nonAM plants exposed to drought alone, but with exposure to combined NaCl and drought stress, stomates of Gi plants remained open 17-22% longer than in nonAM plants. Promotion of stomatal conductance by Gm occurred with exposure to NaCl/drought stress but not with drought alone or with soil leaching before drought. In other instances, however, the addition of salt tended to nullify an AM-induced change in drought response. Our findings confirm that AM fungi can alter host response to drought but do not lend much support to the idea that AM-induced salt resistance might help explain why AM plants can be more resilient to drought stress than their nonAM counterparts.     

Proteomic study of Carissa spinarum in response to combined heat and drought stress

Carissa spinarum is one of the secondary advantage plants grown in dry‐hot valleys in China, which can survive under stress conditions of high temperature and extreme low humidity. Here, we studied the physiological and proteomic changes of C. spinarum in response to 42°C heat stress treatment in combination with drought stress. Dynamic changes in the leaf proteome were analyzed at four time points during the stress treatment and recovery stages. Approximately, 650 protein spots were reproducibly detected in each gel. Forty‐nine spots changed their expression levels upon heat and drought treatment, and 30 proteins were identified by MS and 2‐D Western blot. These proteins were classified into several categories including HSP, photosynthesis‐related protein, RNA‐processing protein and proteins involved in metabolism and energy production. The potential roles of these stress‐responsive proteins are discussed.     

植物对干旱胁迫的分子反应

干旱胁迫是影响植物生长发育的主要因子,渗透保护剂的合成和积累,脱水伤害的修复,自由基清除酶和LEA蛋白基因表达的增量调节能增加植物的耐干旱性。植物在干旱条件下至少有4条信号转导途径,其中2条信号途径是依赖ABA的,另外2条途径是不依赖ABA的,在植物干旱胁迫的信号转导中,双组分的组氨酸激酶可能起渗透感受器的作用,Ca^2 和IP3可能是脱水信号的第2信使,转基因植物是一种评价编码蛋白功能的良好系统。     

Identification and profiling of salinity stress-responsive proteins in Sorghum bicolor seedlings

Sorghum bicolor, a drought tolerant cereal crop, is not only an important food source in the semi arid/arid regions but also a potential model for studying and gaining a better understanding of the molecular mechanisms of drought and salt stress tolerance in cereals. In this study, seeds of a sweet sorghum variety, MN1618, were planted and grown on solid MS growth medium with or without 100mM NaCl. Heat shock protein expression immunoblotting assays demonstrated that this salt treatment induced stress within natural physiological parameters for our experimental material. 2D PAGE in combination with MS/MS proteomics techniques were used to separate, visualise and identify salinity stress responsive proteins in young sorghum leaves. Out of 281 Coomassie stainable spots, 118 showed statistically significant responses (p<0.05) to salt stress treatments. Of the 118 spots, 79 were selected for tandem mass spectrometric identification, owing to their good resolution and abundance levels, and of these, 55 were positively identified. Identified proteins were divided into six functional categories including both known and novel/putative stress responsive proteins. Molecular and physiological functions of some of our proteins of interest are currently under investigation via bioinformatic and molecular biology approaches.     

Patterns of allozyme variation in cultivated and wild Sorghum bicolor

Summary Patterns of allozyme variation were surveyed in collections of cultivated and wild sorghum from Africa, the Middle East, and Asia. Data for 30 isozyme loci from a total of 2067 plants representing 429 accessions were analyzed. Regional levels of genetic diversity in the cultivars are greater in northern and central Africa compared to southern Africa, the Middle East, or Asia. The spatial distribution of individual alleles at the most variable loci was studied by plotting allele frequencies on geographic maps covering the distribution of sorghum. Generally, many of the alleles with frequencies below 0.25 are localized in specific portions of the range and are commonly present in more than one race in that region. Several alleles occur in both wild and cultivated sorghum of one region and are absent from sorghum elsewhere, suggesting local introgression between the wild and cultivated forms. Although the same most common allele was found in the wild and cultivated gene pools at 29 of the 30 loci, phenetic analyses separated the majority of wild collections from the cultivars, indicating that the two gene pools are distinct. Wild sorghum from northeast and central Africa exhibits greater genetic similarities to the cultivars compared to wild sorghum of northwest or southern Africa. This is consistent with the theory that wild sorghum of northeast-central Africa is ancestral to domesticated sorghum. Wild sorghums of race arundinaceum of northwest Africa and race virgatum from Egypt are shown to be genetically distinct from both other forms of wild sorghum and from the cultivars. Suggestions for genetic conservation are presented in light of these data.     

高粱CRY2基因的克隆及其表达分析

从高粱(Sorghum bicolor L．var．R111)幼苗中提取总RNA,利用RT-PCR和cDNA的3′末端的快速扩增方法(3′RACE),第一次克隆了高粱隐花色素2基因(CRY2)的cDNA序列。该序列包括了一个完整的开放阅读框,编码大小为690个氨基酸残基的蛋白质,与水稻、番茄和拟南芥CRY2蛋白质的同源性分别为87％、57％和45．5％。高粱CRY2基因组DNA含有3个内含子和4个外显子。RT-PCR检测结果表明,高粱CRY2基因在根、茎和叶中都有转录。Western blotting结果显示CRY2蛋白在根、茎和叶中表达,并在黑暗中积累,蓝光下降解。高粱CRY2可能在蓝光诱导的幼苗去黄化反应中起作用。     

高梁CRY2基因的克隆及其表达分析

从高粱(Sorghum bicolor L. var.R1ll)幼苗中提取总RNA,利用RT-PCR和cDNA的3'末端的快速扩增方法(3'RACE),第一次克隆了高粱隐花色素2基因(CRY2)的cDNA序列.该序列包括了一个完整的开放阅读框,编码大小为690个氨基酸残基的蛋白质,与水稻、番茄和拟南芥CRY2蛋白质的同源性分别为8 7%、5 7%和45.5%.高粱CRY2基因组DNA含有3个内含子和4个外显子.RT-PCR检测结果表明,高粱CRY2基因在根、茎和叶中都有转录.Western blotting结果显示CRY2蛋白在根、茎和叶中表达,并在黑暗中积累,蓝光下降解.高粱CRY2可能在蓝光诱导的幼苗去黄化反应中起作用.     

沙地云杉生态型对干旱胁迫的生理生态响应

采用干旱胁迫处理不同生态型沙地云杉幼苗,对其主干高生长、丙二醛(MDA)、脯氨酸和叶绿素含量进行了测定．结果表明,沙地云杉幼苗高生长明显受到干旱胁迫的抑制,绿果型沙地云杉在水分条件良好的情况下,高生长迅速,明显高于紫果型沙地云杉．随着干旱胁迫强度的增强,其高生长受抑制的程度高于紫果型沙地云杉．红果型沙地云杉处于二者之间．同时干旱胁迫也影响沙地云杉生长进程,使得其封顶时间提前一个多月．MDA含量无论在叶还是根中呈上升趋势,只是在土壤含水量在10％左右出现轻度胁迫时略有下降;叶片中MDA含量明显高于根中;从不同生态型来看,紫果型沙地云杉体内MDA含量高,绿果型低且具有较大的变化幅度．脯氨酸含量随干旱胁迫强度增加而增加;绿果型沙地云杉根部脯氨酸含量在处理4之前高于另外两种生态型．叶绿素的变化规律,紫果型和红果型沙地云杉基本相同,只是红果型含量略微低于紫果型．其叶绿素a、叶绿素b和总叶绿素含量都是开始逐渐升高;随着胁迫强度增加,叶绿素含量下降;绿果型沙地云杉叶绿素含量总体上都呈下降趋势,只有叶绿素b含量在胁迫初期有所增加．从叶绿素a／b值出现下降趋势,但不同生态型表现不一样．绿果型沙地云杉由最高下降到最低,紫果型沙地云杉下降幅度较小,红果型介于二者之间．表明不同生态型沙地云杉抗旱能力不同,紫果型>红果型>绿果型．     

高粱SBP-box基因家族全基因组鉴定及表达分析

SQUAMOSA PROMOTER BINDING PROTEIN box (SBP-box)基因家族编码一类绿色植物特有的转录因子,其功能涉及作物遗传改良的许多方面,如产量、株型、抗逆性等,具有重要的实际应用价值。本研究通过生物信息学方法从高粱(Sorghum bicolor L.)全基因组中鉴定出18个SBP-box基因,分布于9条染色体上,其中8个基因位于基因组重复区域。系统发育分析表明高粱SBP-box基因家族可分为6个亚家族,其中SbSBP12、SbSBP3和SbSBP15分别与玉米ZmLG1、ZmTGA1和ZmUB2/3直系同源。基于RNA-seq数据的表达分析发现高粱SBP-box基因在花序原基中表达量最高,SbSBP9和SbSBP17为花序原基特异表达基因,SbSBP5、SbSBP8和SbSBP18等基因受外源ABA和PEG胁迫上调表达,表明SBP-box基因可能参与高粱对非生物逆境的响应。本研究为高粱SBP-box家族重要基因的克隆提供了参考,相关基因可作为高粱遗传改良的候选基因。     

Reactions of Egyptian landraces of Hordeum vulgare and Sorghum bicolor to drought stress, evaluated by the OJIP fluorescence transient analysis

Landraces of barley and of Sorghum bicolor from Egypt were evaluated for their tolerance to drought stress (DS) using the OJIP test of the chl fluorescence fast induction curve. Water was withheld from 4-week-old, pot-grown plants for 8–10 days, until the volumetric soil water content decreased from 30 to below 5 vol% and the leaves reached relative water contents of <60 %. The plants were rewatered and recovery measurements were taken 24 h later. Comparative studies of the most sensitive and the most tolerant lines of both cereals, as evaluated by their Performance Indices (PI_abs), revealed a similar behavior in the sensitive lines, i.e., inhibiting effects of DS on PS II connectivity (occurrence of an L band), oxygen evolving complex (occurrence of a K band) and on the J step of the induction curves, associated with an inhibition of electron transport from Q _A to Q _B . These effects persisted or were even enhanced in the rewatered plants, which resulted in similar deviations of spider plots of the OJIP parameters in the sensitive lines of both species. In the most tolerant barley accession, drought effects on “early” events (i.e., L, K bands) were much smaller or negligible, and there was no pronounced effect on the J step. However, distinct increases of the I step occurred, pointing to inhibited electron flow to the intersystem electron carriers and beyond PS I. The most tolerant Sorghum line, in contrast, revealed nearly no effects of the DS and recovery treatment on the fluorescence induction curves and OJIP parameters.     

天山北坡前山带12种植物对干旱胁迫和复水的响应及抗旱性评价

【目的】探讨12种植物对干旱胁迫的动态生理适应性及抗旱能力,筛选适宜在天山北坡前山带进行植被恢复的灌木树种。【方法】以新疆天山北坡前山带种植的刺蔷薇、柠条锦鸡儿、文冠果、山杏、柽柳、梭梭、中亚沙棘、蒙古沙棘（‘新棘1号’、‘新棘2号’、‘新棘3号’、‘新棘4号’、‘新棘5号’）共12种植物为研究对象,观测灌水之后土壤水分的时空分布变化,分析干旱胁迫和复水处理对植物光合气体交换参数、抗逆生理生化指标、叶绿素荧光参数的影响,利用主成分分析法评价12种植物的抗旱能力。【结果】（1）在干旱胁迫发生前后,各植物立地土壤表层（0—20 cm）水分散失最快,中层（20—40 cm）次之,底层（40—60cm）水分散失最慢,而不同植物因其生长策略不同,通过改变其生长情况来适应干旱;（2）随着干旱胁迫的加剧,植物叶片光合生理指标净光合速率、气孔导度、蒸腾速率、胞间二氧化碳浓度、PSⅡ潜在活性、 PSⅡ最大光化学效率、电子传递速率和叶绿素含量降低,而抗逆生理指标超氧化物歧化酶活性、脯氨酸含量、丙二醛含量及非光化学猝灭系数升高,复水之后各指标都有一定程度的恢复,但均未达到初始水平;（3）主成分分析结果显示,净光合速率、叶绿素含量、电子传递效率和丙二醛含量等指标贡献率较大,梭梭、柽柳、刺蔷薇和柠条锦鸡儿抗旱性较强,中亚沙棘、‘新棘5号’、‘新棘1号’和山杏抗旱性次之,而‘新棘4号’、‘新棘3号’、文冠果和‘新棘2号’较弱。【结论】净光合速率、叶绿素含量、电子传递效率和丙二醛含量可作为评价各植物适应干旱胁迫的重要指标;新疆天山北坡前山带区域植被生态恢复过程中应优先考虑梭梭、柽柳、刺蔷薇及柠条锦鸡儿进行建植。