The
15N natural abundance method has been widely used for evaluation of symbiotic N
2-fixation. The method inevitably requires a reference plant that reflects soil derived δ
15N similar to that in a N
2-fixing target plant, for estimating the contribution of fixed N
2. However, it is often difficult to select a suitable reference plant. Recently, an alternative method was proposed using the difference in δ
15N values between shoots and nodulated roots, which did not require a reference plant
per se. Whether this method is applicable to a wide range of N
2-fixing plants having different growth habits and symbiosis types remains to be verified. To test the applicability of this method for perennial plants, we examined the difference in δ
15N values between shoot and nodulated root (Δδ
15N
s-nr), and that between shoot and root (Δδ
15N
s-r) in 6-month-old plants grown in pots with different soil moisture regimes. The relationships between Δδ
15N
s-nr and the percentage of N derived from atmospheric N
2 (%Ndfa) calculated from the conventional
15N natural abundance method, and between Δδ
15N
s-r and %Ndfa were analyzed in N
2-fixing legume
Lespedeza cuneata and N
2-fixing non-legume
Elaeagnus pungens and
Myrica rubra. A close correlation was found between Δδ
15N
s-nr and %Ndfa as well as between Δδ
15N
s-r and %Ndfa in
Lespedeza cuneata, while no correlation was found in N
2-fixing non-legume species. The results indicated that Δδ
15N signatures could be useful for estimating %Ndfa for N
2-fixing perennial legume (
Lespedeza cuneata) in the first growth season but might not be applicable for N
2-fixing actinorhizal plants.
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