Abstract

Alfalfa (Medicago sativa L.) is an important perennial forage, with high nutritional value, which is widely grown in the world. Because of low freezing tolerance, its distribution and production are threatened and limited by winter weather. To understand the complex regulation mechanisms of freezing tolerance in alfalfa, we performed transcriptome sequencing analysis under cold (4 °C) and freezing (-8 °C) stresses. More than 66 million reads were generated, and we identified 5767 transcripts differentially expressed in response to cold and/or freezing stresses. These results showed that these genes were mainly classified as response to stress, transcription regulation, hormone signaling pathway, antioxidant, nodule morphogenesis, etc., implying their important roles in response to cold and freezing stresses. Furthermore, nine CBF transcripts differentially expressed were homologous to CBF genes of Mt-FTQTL6 site, conferring freezing tolerance in M. truncatula, which indicated that a genetic mechanism controlling freezing tolerance was conservative between M. truncatula and M. sativa. In summary, this transcriptome dataset highlighted the gene regulation response to cold and/or freezing stresses in alfalfa, which provides a valuable resource for future identification and functional analysis of candidate genes in determining freezing tolerance.

Highlights

  • Plants are always challenged by various unfavorable environmental conditions, such as low temperatures, water deficit, and salinity

  • Nine C-repeat binding factors (CBF) transcripts differentially expressed were homologous to CBF genes of Mt-FTQTL6 site, conferring freezing tolerance in M. truncatula, which indicated that a genetic mechanism controlling freezing tolerance was conservative between M. truncatula and M. sativa

  • The results showed that alfalfa unigenes had the highest genetic similarity with M. truncatula (65.1%, 49,216/75,551), followed by Cicer arietinum (44.2%, 33,356/75,551), Glycine max (35.0%, 26,464/75,551), Phaseolus vulgaris (29.1%, 21,983/75,551), Lotus japonicus (27.3%, 20,651/75,551), while Arabidopsis was the lowest (3.5%, 2,631/75,551); see Figure 2

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Summary

Introduction

Plants are always challenged by various unfavorable environmental conditions, such as low temperatures, water deficit, and salinity. Among these stresses, low temperatures are one of the major factors, limiting plant growth, development, and distribution. In order to adapt to low temperature conditions, plants have employed numerous regulation mechanisms to survive through cold and/or freezing stresses (Knight and Knight, 2012). Important progress have revealed that prior exposure to non-freezing low temperatures improved freezing tolerance of a plant, which is known as cold acclimation (Thomashow, 1999). In plants, including Arabidopsis, the molecular regulation mechanisms of cold acclimation and acquired freezing tolerance have been extensively investigated. Many reports have showed that Crepeat (CRT) binding factors (CBF), known as dehydration responsive elements binding factors (DREB), have played fundamental roles in plant adaption to low tempera-

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