Salt-tolerant and -sensitive alfalfa (<i>Medicago sativa</i>) cultivars have large variations in defense responses to the lepidopteran insect <i>Spodoptera litura</i> under normal and salt stress condition

<div><p>In nature, plants are often exposed to multiple stress factors at the same time. Yet, little is known about how plants modulate their physiology to counteract simultaneous abiotic and biotic stresses, such as soil salinity and insect herbivory. In this study, insect performance bioassays, phytohormone measurements, quantification of transcripts, and protein determination were employed to study the phenotypic variations of two alfalfa (<i>Medicago sativa</i>) cultivars in response to insect <i>Spodoptera litura</i> feeding under normal and salt stress condition. When being cultivated in normal soil, the salt-tolerant alfalfa cultivar Zhongmu-1 exhibited lower insect resistance than did the salt-sensitive cultivar Xinjiang Daye. Under salinity stress, the defense responses of Xinjiang Daye were repressed, whereas Zhongmu-1 did not show changes in resistance levels. It is likely that salinity influenced the resistance of Xinjiang Daye through suppressing the accumulation of jasmonic acid-isoleucine (JA-Ile), which is the bioactive hormone inducing herbivore defense responses, leading to attenuated trypsin proteinase inhibitor (TPI) activity. Furthermore, exogenous ABA supplementation suppressed the insect herbivory-induced JA/JA-Ile accumulation and levels of <i>JAR1</i> (<i>jasmonate resistant 1</i>) and TPI, and further decreased the resistance of Xinjiang Daye, whereas Zhongmu-1 showed very little response to the increased ABA level. We propose a mechanism, in which high levels of abscisic acid induced by salt treatment may affect the expression levels of <i>JAR1</i> and consequently decrease JA-Ile accumulation and thus partly suppress the defense of Xinjiang Daye against insects under salt stress. This study provides new insight into the mechanism by which alfalfa responds to concurrent abiotic and biotic stresses.</p></div

JA and JA-Ile contents in two alfalfa cultivars in response to <i>S</i>. <i>litura</i> feeding.

<p>Zhongmu-1 and Xinjiang Daye were treated with W+OS, and samples were harvested at 0, 0.5, 1, 1.5 and 3 h, and the JA (A) and JA-Ile (B) levels were determined (n = 5). Asterisks indicate significances between two cultivars with the same treatment and time point (Unpaired <i>t</i>-test; *, P < 0.05; ***, P < 0.001).</p

Insect performance on seven different alfalfa cultivars and a two-way choice assay on Zhongmu-1 and Xinjiang Daye.

<p>(A) Larval masses of <i>S</i>. <i>litura</i> feeding on seven different cultivars of alfalfa (n = 150). (B) A photograph and (C) a bar-graph showing the two-way choice test assessing the leaf tissue consumption of <i>S</i>. <i>litura</i> larvae on Zhongmu-1 and Xinjiang Daye. Different lowercase letters indicate significant differences among different cultivars (Tukey HSD test; P < 0.05); asterisks indicate significantly different levels between two cultivars (paired <i>t</i>-test; ***, P < 0.001).</p

<i>S</i>. <i>litura</i> growth and ABA contents under normal and salt-stress condition.

<p>Zhongmu-1 and Xinjiang Daye were irrigated with 250 mM NaCl or water and thereafter cultivated for a week. (A) Masses of <i>S</i>. <i>litura</i> feeding on these plants (n = 150). (B) ABA levels in Zhongmu-1 and Xinjiang Daye, 1 h after W+OS treatment (untreated plants served as controls; n = 5). Different lowercase letters represent significant differences among the combinations of abiotic stresses and cultivars. Different uppercase letters indicate significant differences between biotic stresses within the same cultivar and abiotic treatment (Tukey HSD test; P < 0.05).</p

Additional file 1: of Comparative analysis of alfalfa (Medicago sativa L.) leaf transcriptomes reveals genotype-specific salt tolerance mechanisms

This PDF contains all of the additional material (Figure S1–S5) associated with the manuscript. Figure numbers and titles are listed below: Figure S1. The survival rates of seven alfalfa cultivars. Figure S2. The growth phenotypes of XJ and ZM cultivars. Figure S3. Distribution of the lengths of the assembled unigenes in the transcriptomes of alfalfa leaf tissue. Figure S4. The GO and KEGG annotations of DEGs between XJ and ZM. Figure S5. The GO and KEGG annotations of the salt-responsive genes compared between XJ and ZM. (PDF 1351 kb

Additional file 2: of Comparative analysis of alfalfa (Medicago sativa L.) leaf transcriptomes reveals genotype-specific salt tolerance mechanisms

This excel file contains all of the additional Tables (Table S1-S9) associated with the manuscript. Each Table is in a different tab. Table numbers and titles are listed below: Table S1. Summary of clean reads and mapping ratio. Table S2. Number of annotated unigenes in alfalfa. Table S3. Annotations of DEGs in two cultivars of alfalfa. Table S4. The DEGs involved in ROS homeostasis in two cultivars. Table S5. The DEGs involved in Ca2+ signaling in two cultivars. Table S6. The DEGs involved in phytohormone biosynthesis in two cultivars. Table S7. The DEGs involved in ion transport in two cultivars. Table S8. The DEGs involved in transcription factor in two cultivars. Table S9. The photosynthesis-related DEGs in two cultivars. (XLSX 6789 kb