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Genome-Wide Survey and Development of Novel SSR Markers in Erianthus fulvus
Scientia Agricultura Sinica 2025, 58(5): 851-863
Published: 01 March 2025
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【Objective】

Erianthus fulvus, serving as a crucial wild resource for sugarcane, is capable of enhancing the stress tolerance and yield of varieties. In order to utilize E. fulvus for sugarcane breeding, it is important to systematically identify and develop simple sequence repeat (SSR) loci in the E. fulvus genome, screen for polymorphic SSR markers, analyse the genetic diversity characteristics of E. fulvus resources and then develop SSR markers associated with important traits.

【Method】

Using the SSRminer module in the software TBtools, a comprehensive exploration of SSR loci was conducted on the diploid E. fulvus whole genome sequence. The obtained data were statistically analyzed to reveal their distribution patterns and regularities within the genome. The Batch Target Region Primer Design function was employed for batch designing SSR primers, and the specificity of the primers was evaluated using the Primer check tool. To comparethe SSR polymorphism betweenE. fulvus and sugarcane, amplification experiments were performed on 50 pairs of randomly synthesized SSR primers and 14 pairs of SSR primers sourced from sugarcane across 6 E. fulvus germplasms.

【Result】

A total of 152 707 SSR loci, which were distributed on E. fulvus genome with an average density of 5.64 kb/locus, were identified. The majority were located in intergenic regions. In terms of SSR type distribution, mononucleotide, dinucleotide, and trinucleotide had the highest density. Dinucleotide SSR types exhibited the greatest variation in motif repeat numbers, while pentanucleotide motif repeat variations were the least. Across the entire genome, 883 distinct SSR motif repeat types were identified, with A/T and AT/TA being the most abundant. A total of 144 692 pairs of SSR primers, of which 85 025 pairs exhibited high specificity, were designed. These specific primers displayed a distribution characteristic of dense ends and sparse middles on the genome. Amplification experiments showed that 42 out of the 50 randomly synthesized SSR primer pairs yielded stable and clear bands in E. fulvus, with 32 exhibiting polymorphisms, yielding a polymorphism rate of 64.0%. In contrast to the 14 sugarcane SSR primers, the E. fulvus SSR primers demonstrated superior amplification efficacy and greater polymorphism. After screening, 16 pairs of SSR primers with good polymorphism and clear amplification bands were determined from the 32 effective SSR primer pairs. These 16 pairs of primers amplified a total of 72 bands, with polymorphism information content (PIC) ranging from 0.63 to 0.83, and an average PIC value of 0.74, indicating their effectiveness and practicality in polymorphism analysis and molecular marker research of E. fulvus germplasm resources.

【Conclusion】

This study comprehensively identified SSR loci in the E. fulvus genome, revealing the high abundance and diversity of SSR distribution features. Sixteen pairs of highly specific and polymorphic SSR primers were successfully screened.

Issue
Application of Trehalose Enhances Drought Resistance in Sugarcane Seedlings and Promotes Plant Growth
Scientia Agricultura Sinica 2023, 56(21): 4208-4218
Published: 01 November 2023
Abstract PDF (1.7 MB) Collect
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【Objective】

This study aims to investigate whether application of trehalose could mitigate the adverse effects of drought stress by enhancing the defense response in sugarcane (Saccharum spp. hybrid), and to provide a theoretical basis for stable high yield of sugarcane under drought conditions.

【Method】

The present investigation was conducted to assess ameliorative effects of adding trehalose on the growth, malondialdehyde (MDA) content, and antioxidant responses of two sugarcane cultivars ROC22 and YZ05-51 under the treatment of 30% PEG6000 and 12% PEG6000. In the preliminary experiment, four treatment concentrations of trehalose were set at 0, 10, 100 and 200 mg·L-1. The dry weight, fresh weight and adventitious roots of tissue-cultured sugarcane seedlings were measured at 9 d after treatment to clarify the optimal concentration of trehalose in promoting sugarcane growth under drought stress. Then, the MDA content and the activity of antioxidant enzyme peroxidase (POD) and superoxide dismutase (SOD) were determined under control, drought stress and drought stress combined with optimal trehalose treatment. Finally, the expression level of drought-resistance genes ScTPS1, ScSnRK2.3, ScSnRK2.4 and ScDREB2b-1 was quantified by qRT-PCR at 0, 12, 24 and 48 h in YZ05-51 after exposed to 30% PEG6000 and 30% PEG6000 adding with optimal trehalose concentration, respectively.

【Result】

Application of trehalose could alleviate the drought-induced decrease of fresh weight and dry weight of two sugarcane cultivars, and the growth recovery of plantlet was better in 100 mg·L-1 trehalose group than that in 10 and 200 mg·L-1 trehalose groups. Under 30% PEG6000 stress, the MDA content was notably increased, and a considerable improvement was recorded in the activity of the antioxidant enzymes POD and SOD. Adding trehalose significantly reduced the content of drought-induced MDA, and enhanced the activity of POD and SOD, respectively. However, external trehalose had little effect on the MDA content and antioxidant enzyme activity of tissue-cultured sugarcane seedlings under 12% PEG6000 stress. Compared with drought controls, the expression of drought-resistance genes ScTPS1, ScSnRK2.3, ScSnRK2.4 and ScDREB2b-1 was up-regulated in the trehalose-treated plantlet.

【Conclusion】

Application of trehalose can alleviate the negative impact of drought on the growth of sugarcane seedlings, and promote adventitious root growth. External addition of trehalose may reduce the oxidative toxicity caused by drought stress by increasing the activity of antioxidant enzymes POD and SOD. Meanwhile, application of trehalose induces the expression of drought-resistance genes ScTPS1, ScSnRK2.3, ScSnRK2.4, and ScDREB2b-1, indicating that applying trehalose can improve the drought resistance of sugarcane. The results will provide a reference for developing drought resistance strategies in sugarcane breeding and productive practice.

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