The impact of phytochemical, morpho-physiological, and biochemical changes of Lallemantia royleana (Benth.) on drought tolerance

Figures & data

Figure 1. Monthly change of minimum and maximum temperatures and rainfall rates in 2015 and 2016.

Table 1. Specifications of Balangu ecotypes collection areas.

Ecotype	Geographical coordinates	Elevation (meter)	Region climate (Köppen-Geiger classification)
Kalat	59° 45ʹ N and 36° 59ʹ E	926	Csb
Kondor	51° 07ʹ N and 35° 51ʹ E	1979	BSk
Jupar	57° 15ʹ N and 29° 57ʹ E	2434	BWk
Zakheh	47° 06ʹ N and 35° 45ʹ E	1965	DSa

Table 2. Soil physicochemical properties of experimental farm at 2015 and 2016.

Year	Clay (%)	Silt (%)	Sand (%)	Texture	N (%)	P (ppm)	K (ppm)	OC (%)	EC (ds m ^−1)	pH
2015	20	33	47	loamy	0.09	8.9	245	0.59	3.8	7.8
2016	18	31	51	loamy	0.11	9.4	267	0.67	3.3	7.6

Figure 2. The drought impact on the dry weight (a), seed yield (b), leaf numbers (c), plant height (d), and HI (e) of various Lallemantia royaleana ecotypes. Values are means (n = 6). Circle = Kalat ecotype, triangle = Kondor ecotype, rectangle = Jupar ecotype, and diamond = Zakheh ecotype.

Figure 3. The drought impact on RWC of various Lallemantia royaleana ecotypes. Values are means (n = 3). Means with the same letter have no significant difference at p ≤ 0.05.

Table 3. Mean comparison of Lallemantia royleana growth features at different drought stress and ecotype.

Drought (atm)	Ecotype	Seed yield (kg ha^−1)		Plant dry weight (ton ha^−1)	HI		Plant height (cm)		Leaf number
		2015	2016		2015	2016	2015	2016
Control (- 0.5)	Kalat	1289 ^a	1166 ^b	1.99 ^d	39.1 ^a	37.4 ^a	34.8 ^b	34.1 ^ab	217.7 ^b
	Kondor	1332 ^a	1293 ^a	2.80 ^b	32.0 ^bc	32.6 ^bc	34.8 ^b	33.7 ^ab	199.7 ^b
	Jupar	466 ^f	455 ^f	1.54 ^ef	24.8 ^de	22.0 ^e	34.7 ^b	33.2 ^bc	181.1 ^c
	Zakheh	628 ^e	606 ^e	3.42 ^a	16.2 ^fg	14.6 ^f	40.7 ^a	35.8 ^a	240 ^a
Mild (- 3.5)	Kalat	843 ^c	843 ^d	1.73 ^de	30.5 ^bc	35.9 ^ab	29.9 ^bcd	30.0 ^def	147.7 ^de
	Kondor	1004 ^b	1068 ^c	2.01 ^d	33.7 ^ab	34.5 ^ab	30.8 ^bc	32.4 ^bcd	157.7 ^d
	Jupar	420 ^fg	423 ^fg	1.08 ^gh	29.3 ^bcd	27.5 ^d	30.8 ^bc	30.1 ^def	130.4 ^ef
	Zakheh	437 ^f	447 ^f	2.33 ^c	17.2 ^fg	15.2 ^f	30.9 ^bc	30.9 ^cde	153.9 ^d
Moderate (- 6.5)	Kalat	298 ^h	360 ^gh	1.30 ^fg	19.5 ^ef	21.2 ^e	28.5 ^cde	26.9 ^g	78.1 ^i
	Kondor	713 ^d	835 ^d	1.26 ^fg	38.1 ^a	38.3 ^a	29.3 ^cde	30.2 ^def	117.2 ^fg
	Jupar	401 ^fg	418 ^fgh	1.05 ^gh	28.4 ^bcd	28.0 ^d	25.9 ^cdef	29.8 ^ef	105 ^gh
	Zakheh	152 ^i	152 ^ij	1.15 ^gh	13.4 ^g	10.5 ^g	25.3 ^def	26.9 ^g	88.4 ^hi
Severe (−9.5)	Kalat	0.0 ^j	180 ^i	0.45 ^i	0.0 ^h	16.8 ^f	0.0 ^g	20.8 ^h	23.3 ^j
	Kondor	391 ^fg	473 ^f	1.08 ^gh	27.9 ^cd	29.2 ^cd	24.5 ^ef	28.2 ^fg	83.8 ^i
	Jupar	341 ^gh	345 ^h	0.90 ^h	28.4 ^bcd	26.9 ^d	23.0 ^f	27.8 ^fg	84.5 ^i
	Zakheh	0.0 ^j	86.7 ^j	0.43 ^i	0.0 ^h	9.1 ^g	0.0 ^g	20.3 ^h	22.8 ^j

Means in each column with the same letters have no significant difference at p ≤ 0.05.

Figure 4. The effect of drought stress on the seed oil content of various ecotypes (a) and omega −6 content (b), and the content change of omega-6 (c) and the α-linolenic acid (c) in different ecotypes in 2015 and 2016. Values are means for seed oil content (n = 6), α-linolenic acid and omega-6 content (n = 3). Different alphabet characters indicate significant differences (p ≤ 0.05), determined by Duncan test.

Figure 5. Change of protein (a), phenol (c), and soluble carbohydrate (d) contents in response to drought stresses in the various ecotypes. Values are means (n = 6).

Table 4. Mean comparison of Lallemantia reoyleana biochemical characteristics at different drought stress and ecotype.

Drought (atm)	Ecotype	MC (%)		Phenol (mg g^−1 FW)		APX (mM mg^−1 Pro min^−1)		CAT (mM mg^−1 Pro min^−1)		SOD (Unit g^−1 FW)		POX (mM mg^−1 Pro min^−1)
		2015	2016	2015	2016	2015	2016	2015	2016	2015	2016	2015	2016
Control (- 0.5)	Kalat	7.6 ^g	9.7 ^g	13.4 ^i	19.8 ^ghi	11.4 ^ef	8.4 ^h	4.0 ^ef	4.1 ^f	66.7 ^de	59.5 ^fg	0.9 ^efg	1.1 ^ghi
	Kondor	12.8 ^cd	13.6 ^de	18.7 ^h	23.1 ^efg	14.6 ^e	17.6 ^f	1.2 ^gh	1.0 ^gh	49.3 ^f	60.0 ^fg	0.6 ^h	0.9 ^i
	Jupar	11.7 ^de	12.8 ^ef	26.5 ^e	19.2 ^hi	5.8 ^g	7.6 ^h	0.6 ^gh	0.6 ^h	47.9 ^f	60.3 ^fg	1.3 ^b	1.4 ^cde
	Zakheh	10.2 ^ef	11.4 ^f	6.2 ^j	16.4 ^ij	9.8 ^efg	11.8 ^h	1.6 ^g	1.3 ^gh	50.9 ^f	56.6 ^g	0.9 ^fg	1.0 ^hi
Mild (- 3.5)	Kalat	8.9 ^fg	11.4 ^f	37.8 ^b	35.7 ^a	12.0 ^ef	13.6 ^fg	8.6 ^c	11.3 ^d	79.3 ^cd	65.9 ^ef	1.1 ^cd	1.3 ^efg
	Kondor	18.3 ^b	19.1 ^b	18.5 ^h	24.3 ^ef	22.4 ^d	26.9 ^e	1.5 ^g	3.0 ^fgh	50.3 ^f	75.0 ^d	0.9 ^fg	1.0 ^hi
	Jupar	11.4 ^de	12.8 ^ef	24.8 ^f	26.4 ^de	9.5 ^fg	11.4 ^gh	2.9 ^f	3.5 ^fg	50.1 ^f	62.7 ^efg	1.3 ^bc	1.3 ^def
	Zakheh	11.1 ^e	12.6 ^ef	29.4 ^c	23.6 ^ef	26.2 ^bcd	31.4 ^de	4.9 ^e	8.5 ^e	53.5 ^ef	59.3 ^fg	1.2 ^bcd	1.3 ^ef
Moderate (- 6.5)	Kalat	10.7 ^e	13.0 ^e	28.4 ^cd	22.1 ^fgh	12.5 ^ef	14.5 ^fg	14.5 ^a	17.2 ^c	135.7 ^a	76.7 ^d	1.7 ^a	1.6 ^c
	Kondor	20.4 ^a	22.0 ^a	22.8 ^g	22.1 ^fgh	24.8 ^cd	29.7 ^de	6.5 ^d	11.0 ^d	90.5 ^bc	77.6 ^cd	1.2 ^bc	1.3 ^def
	Jupar	11.1 ^e	13.0 ^e	24.3 ^f	29.6 ^bcd	11.3 ^ef	13.6 ^fg	6.8 ^d	7.6 ^e	67.4 ^de	78.6 ^cd	1.0 ^def	1.2 ^fgh
	Zakheh	13.9 ^c	15.9 ^c	45.0 ^a	31.7 ^bc	31.2 ^b	36.7 ^c	8.6 ^c	19.6 ^c	101.9 ^b	67.7 ^e	1.6 ^a	1.5 ^cd
Severe (−9.5)	Kalat	0 ^h	15.1 ^cd	0.0 ^k	12.2 ^k	0.0 ^h	15.0 ^fg	0.0 ^h	30.0 ^ab	0.0 ^g	86.0 ^b	0.0 ^i	1.9 ^b
	Kondor	10.2 ^ef	13.6 ^de	27.8 ^de	28.4 ^cd	58.4 ^a	70.1 ^a	12.6 ^b	28.9 ^b	86.6 ^c	106.5 ^a	1.1 ^cde	1.8 ^b
	Jupar	10.9 ^e	12.4 ^ef	37.7 ^b	33.2 ^ab	28.3 ^bc	34.0 ^cd	4.4 ^e	9.3 ^de	40.1 ^f	84.2 ^bc	0.7 ^gh	0.9 ^i
	Zakheh	0 ^h	18.1 ^b	0.0 ^k	13.8 ^jk	0.0 ^h	45.7 ^b	0.0 ^h	32.5 ^a	0.0 ^g	79.3 ^bcd	0.0 i	2.1 ^a

Means in each column with the same letters have no significant difference at p ≤ 0.05.

Figure 6. Antioxidant enzyme activity response to drought stress in the various ecotypes. a: CAT activity; b: APX activity; c: SOD activity; d: POD activity. Values are means (n = 6). Circle = Kalat ecotype, triangle = Kondor ecotype, rectangle = Jupar ecotype, and diamond = Zakheh ecotype.

Figure 7. Photosynthesis pigments content response to drought stress in the various ecotypes. Values are means(n = 6). Circle = Kalat ecotype, triangle = Kondor ecotype, rectangle = Jupar ecotype, and diamond = Zakheh ecotype.

Table 5. Mean comparison of Lallemantia reoyleana pigments at different drought stress and ecotype.

Drought (atm)	Ecotype	Cha (mg g^−1 FW)		Chb (mg g^−1 FW)		ChT (mg g^−1 FW)		Car (mg g^−1 FW)
		2015	2016	2015	2016	2015	2016
Control (- 0.5)	Kalat	0.84 ^bcd	0.72 ^c	0.54 ^bc	0.52 ^c	1.18 ^cd	1.28 ^c	69.2 ^c
	Kondor	0.86 ^b	0.91 ^a	0.70 ^a	0.65 ^a	1.41 ^b	1.60 ^a	43.5 ^e
	Jupar	0.95 ^a	0.89 ^a	0.70 ^a	0.63 ^a	1.49 ^ab	1.56 ^a	25.1 ^g
	Zakheh	0.85 ^bc	0.75 ^bc	0.53 ^c	0.53 ^bc	1.23 ^c	1.33 ^bc	65.5 ^cd
Mild (- 3.5)	Kalat	0.78 ^cde	0.68 ^c	0.47 ^cd	0.49 ^c	1.12 ^cd	1.22 ^c	94.7 ^a
	Kondor	0.76 ^def	0.84 ^ab	0.61 ^b	0.60 ^ab	1.15 ^cd	1.48 ^ab	62.1 ^d
	Jupar	0.79 ^bcd	0.69 ^c	0.48 ^cd	0.50 ^c	1.13 ^cd	1.24 ^c	63.3 ^cd
	Zakheh	0.69 ^f	0.53 d	0.34 ^e	0.39 ^d	0.82 ^ef	0.98 ^d	67.3 ^cd
Moderate (- 6.5)	Kalat	0.14 ^i	0.27 ^e	0.21 ^f	0.21 ^e	0.32 ^g	0.57 ^e	36.3 ^f
	Kondor	0.47 ^h	0.70 ^c	0.54 ^bc	0.50 ^c	1.03 ^d	1.25 ^c	76.9 ^b
	Jupar	0.74 ^ef	0.67 ^c	0.49 ^cd	0.48 ^c	1.09 ^d	1.20 ^c	95.1 ^a
	Zakheh	0.54 ^g	0.51 ^d	0.43 ^d	0.37 ^d	0.85 ^e	0.95 ^d	89.1 ^a
Severe (- 9.5)	Kalat	0.00 ^j	0.23 ^e	0.00 ^g	0.18 ^e	0.00 ^h	0.49 ^e	16.3 ^h
	Kondor	0.12 ^i	0.51 ^d	0.43 ^d	0.37 ^d	0.81 ^ef	0.94 ^d	91.8 ^a
	Jupar	0.44 ^h	0.45 ^d	0.42 ^d	0.33 ^d	0.71 ^f	0.85 ^d	49.6 ^e
	Zakheh	0.00 ^j	0.29 ^e	0.00 ^g	0.19 ^e	0.00 ^h	0.56 ^e	16.6 ^h

Means in each column with the same letters have no significant difference at p ≤ 0.05.

Figure 8. Heatmap hierarchical clustering and biplot PCA of morpho-physiological and biochemical features of various Lallemantia royleana ecotypes under drought stress. Z-score is shown the values of standardized means for each row from low (blue) to high (red).

Table 6. Pearson correlation coefficients of measured traits under various irrigation regimes.

	Chla	Chlb	ChlT	Car	PH	LN	PDW	RWC	SY	HI	phenol	proline	protein	APX	POX	SOD	CAT	MC	SC	oil	αLA
Chlb	0.91**
ChlT	0.9**	0.99**
Car	0.34	0.43	0.46
PH	0.85**	0.86**	0.85**	0.45
LN	0.88**	0.81**	0.81**	0.26	0.91**
PDW	0.69**	0.62**	0.62**	0.2	0.79**	0.89**
RWC	0.87**	0.81**	0.8**	0.19	0.89**	0.97**	0.85**
SY	0.67**	0.72**	0.69**	0.26	0.68**	0.76**	0.66**	0.8**
HI	0.57*	0.69**	0.66**	0.48	0.61*	0.49	0.25	0.52*	0.78**
phenol	0.22	0.32	0.29	0.63**	0.34	0.02	−0.05	0.07	0.02	0.36
proline	−0.39	−0.06	−0.06	0.39	−0.15	−0.45	−0.43	−0.48	−0.23	0.22	0.45
protein	−0.23	−0.17	−0.19	0.18	0.11	−0.16	−0.15	−0.04	−0.11	0.14	0.29	0.30
APX	−0.32	−0.02	0.004	0.39	−0.12	−0.31	−0.19	−0.36	−0.16	0.04	0.37	0.82**	−0.04
POX	−0.22	−0.09	−0.08	0.26	0.05	−0.24	−0.27	−0.14	−0.38	−0.1	0.43	0.36	0.65**	0.29
SOD	−0.18	0.07	0.04	0.53	0.22	−0.14	−0.11	−0.06	−0.001	0.32	0.58*	0.68**	0.76**	0.46	0.71**
CAT	−0.88**	−0.71**	−0.67**	0.02	−0.69**	−0.80**	−0.64**	−0.77**	−0.59*	−0.42	0.05	0.56*	0.34	0.54*	0.47	0.45
MC	0.19	0.27	0.22	0.28	0.25	0.06	0.03	0.03	0.25	0.44	0.18	0.29	0.08	0.24	0.19	0.35	−0.12
SC	−0.15	0.01	−0.02	0.49*	0.06	−0.31	−0.36	−0.28	−0.18	0.33	0.73**	0.74**	0.61*	0.38	0.39	0.72**	0.29	0.21
oil	0.43	0.32	0.32	−0.05	0.46	0.62**	0.51*	0.71**	0.64**	0.36	−0.18	−0.57*	0.17	−0.59*	−0.16	−0.08	−0.41	−0.27	−0.34
αLA	0.56*	0.42	0.45	0.23	0.51*	0.59*	0.62**	0.55*	0.36	0.05	0.01	−0.41	−0.28	0.001	0.015	−0.2	−0.48	0.19	−0.42	0.05
Omega-6	0.65**	0.6*	0.58*	−0.04	0.63**	0.71**	0.55*	0.81**	0.69**	0.54*	−0.02	−0.46*	0.15	−0.59*	−0.17	−0.05	−0.62**	−0.09	−0.18	0.87	0.03

*: significant at p ≤ 0.05; **: significant at p ≤ 0.01