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Small Auxin Up RNAs influence the distribution of indole-3-acetic acid and play a potential role in increasing seed size in Euryale ferox Salisb.
Huang Z
,
Bao K
,
Jing Z
,
Wang Q
,
Duan H
,
Zhu Y
,
Zhang S
,
Wu Q
.
Abstract
BACKGROUND: Aquatic Euryale ferox Salisb. is an economically important crop in China and India. Unfortunately, low yield limitations seriously hinder market growth. Unveiling the control of seed size is of remarkable importance in improvement of crops. Here, we generated a new hybrid line (HL) with larger seeds by crossing South Gordon Euryale and North Gordon Euryale (WT) which hasn't been reported before. However, the functional genes and molecular mechanisms controlling the seed size in Euryale ferox Salisb. remain unclear. In this study, we focused on the differentially expressed genes in the auxin signal transduction pathway during fruit development between HL and WT to explore candidate regulatory genes participated in regulating seed size.
RESULTS: Both concentration and localization of indole-3-acetic acid (IAA) at two growth stages of fruits of WT and HL were detected by LC-MS and immunofluorescence. Although IAA content between the two lines did not differ, IAA distribution was significantly different. To elucidate the mechanism and to seek the key genes underlying this difference, RNA-seq was performed on young fruits at the two selected growth stages, and differentially expressed genes related to the auxin transduction pathway were selected for further analysis.
CONCLUSION: Hybrid Euryale ferox Salisb. expressed significant heterosis, resulting in non-prickly, thin-coated, large seeds, which accounted for the significantly larger yield of HL than that of WT. Our study indicated that Small Auxin Up RNAs (SAURs) -mediated localization of IAA regulates seed size in Euryale ferox Salisb. We found that some SAURs may act as a positive mediator of the auxin transduction pathway, thereby contributing to the observed heterosis.
Fig. 1. Appearance of WT and SE. The WT has prickly leaves (a), flowers (d) and fruits (g), whereas the SE generates non-prickly leaves (b), flowers (e) and fruits (h). The HL generation has non-pricky leaves (c), flowers (f) and fruits (i) similar to that of SE
Fig. 2. The seed size of WT and HL was measured. The obviously differences between WT (a) and HL (b) were observed in size of seeds, kernels and coats (bar = 1 cm). And (c) length of both kernel and coat in seed of HL are significantly larger than in WT
Fig. 3. On the 10th week, the length of fruits at different stages was measured. Fruits in HL (top of the Fig. 3) grew significantly faster than in WT at stage III among four stages. * P < 0.05, compared to WT (bar = 1 cm)
Fig. 4. Concentration and distribution of IAA in fruits at stage III were detected by LC-MS and immunofluorescence. (a) Fruits in HL and WT did not show significant difference in concentration of IAA at stage II, while (b) the localization of IAA (red) in WT was partly distributed in prick. S3, stage III. (c) Negative control
Fig. 5. Functional annotation of assembled sequences based on GO categorization. 44,363 unigenes were grouped into “cellular component”, “molecular function”, and “biological process”
Fig. 6. DEGs in the IAA signal transduction pathway between HL and WT at stage II and III. (a) DEGs of HL and WT involved in auxin transduction pathway were marked with red, and (b) the relative expression of ARFs, AUX/IAAs, GH3s, SAURs were analyzed. S2, stage II; S3, stage III
Fig. 7. Relative expression levels of 10 candidates were detected by RT-qPCR to verify the results from RNA-seq. Most candidates in RT-qPCR showed similar expression pattern compared to RNA-seq. n = 4–6, * P < 0.05, ** P < 0.01, *** P < 0.001, compared to WT
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