Differential expression of expressed sequence tags in alfalfa roots under aluminum stress

Abstract

To gain a better understanding of differentially expressed sequence tags (ESTs) for aluminum (Al) tolerance and to investigate the molecular mechanisms of Al toxicity, cDNA subtraction libraries were generated from Al-stressed roots of alfalfa (Medicago sativa L.) compared with no Al-stressed ones, employing suppression subtractive hybridization. Using differential screening technique in which the probes were labeled with DIG, we identified 45 non-redundant ESTs in Al-stressed alfalfa root tips with significantly altered expression. Among the up-regulated ESTs, we have found genes encoding identified proteins, including malate dehydrogenase, 6-phosphogluconate dehydrogenase, peroxidase, and an ABC transporter, while the down-regulate genes included ATPase, secretory carrier membrane protein 2, pectinesterase inhibitor. In addition, two novel ESTs, EW678752 and EY976957, up-and down-regulated by Al stress were sequenced. Analyzed by real-time PCR, the expressions of EST EW678718, EW678739, EY976969 and EW678728, which encode for ABC transporter, malate dehydrogenase, peroxidase and 6-phosphogluconate dehydrogenase correspondingly, increased 1.64-, 2.75-, 3.27-and 6.54-folds, respectively, and the expression of EY976957 encoding for ATPase decreased 3.27 folds. The expression of EST EW678752 increased 34.54-fold, while that of EY976957 decreased 16.68 folds. It suggested that the two novel ESTs maybe play a significant role in the aluminum tolerance of alfalfa.

Mechanisms on Boron-Induced Alleviation of Aluminum-Toxicity in Citrus grandis Seedlings at a Transcriptional Level Revealed by cDNA-AFLP Analysis

“…We found that the expression of probable pectate lyase 8-like (TDF #124-1) in roots was down-regulated by Al-toxicity ( Table 1 ), as previously obtained on Al-treated roots of aspen ( Populus tremula ) [ 24 ], indicating that pectate lyases might play a role in plant Al-tolerance. By contrast, the expression of gene encoding probable pectinesterase/pectinesterase inhibitor 61-like (TDF #17-1) was detected only in 2.5 μM B + 1.2 mM Al-treated roots ( Table 1 ), which disagrees with the previous report that Al downregulated alfalfa root expression of pectinesterase inhibitor gene [ 22 ].…”

Identification of sequences expressed during compatible black pepper—Fusarium solani f. sp. piperis interaction

“…piperis infection by SSH in order to improve our understanding about this plant-pathogen interaction, since this crop production is largely affected by root rot disease in the Amazon region. Our subtraction process was validated by PCR amplification of constitutively expressed actin gene that was drastically reduced in subtracted sample appearing first at 33 cycles, as reported by others (Alex et al 2008;Fan et al 2010).…”

“…piperis infection by suppression subtractive hybridization (SSH) (Diatchenko et al 1996), which has been used with success to identify differentially expressed genes (Aslam et al 2010;Fan et al 2010;Gor et al 2010). Our results revealed the identification of sequences coding for putative proteins related to oxidative burst and defense response processes, such as superoxide dismutase, cytochrome p450, cysteine proteinase inhibitor, and pathogenesis-related protein 4.…”

Envolvimento de enzimas NADP-desidrogenases na tolerância ao alumínio em plântulas de soja

“…al., 2021). Além disso, o aumento na atividade da 6PGDH em situações de estresse abiótico, pode fornecer intermediários como açúcares e aminoácidos para as plantas (Fan et. al., 2011;Kaur e Asthir, 2015).…”

“…al, 2018). A atividade da enzima 6PGDH aumentou após exposição a ao alumínio em plantas de alfafa, fornecendo intermediários, como açúcares e aminoácidos para a planta (Fan et. al., 2011).…”