The remarkable regeneration capability of plant tissues or organs under culture conditions has underlain an extensive practice for decades. The initial step in plant in vitro regeneration often involves the induction of a pluripotent cell mass termed callus, which is driven by the phytohormone auxin and occurs via a root development pathway. However, the key molecules governing callus formation remain unknown. Here we demonstrate that Arabidopsis LATERAL ORGAN BOUNDARIES DOMAIN (LBD)/ASYMMETRIC LEAVES2-LIKE (ASL) transcription factors are involved in the control of callus formation program. The four LBD genes downstream of AUXIN RESPONSE FACTORs (ARFs), LBD16, LBD17, LBD18 and LBD29, are rapidly and dramatically induced by callus-inducing medium (CIM) in multiple organs. Ectopic expression of each of the four LBD genes in Arabidopsis is sufficient to trigger spontaneous callus formation without exogenous phytohormones, whereas suppression of LBD function inhibits the callus formation induced by CIM. Moreover, the callus triggered by LBD resembles that induced by CIM by characteristics of ectopically activated root meristem genes and efficient regeneration capacity. These findings define LBD transcription factors as key regulators in the callus induction process, thereby establishing a molecular link between auxin signaling and the plant regeneration program.
The plant hormone auxin plays a critical role in regulating various aspects of plant growth and development, and the spatial accumulation of auxin within organs, which is primarily attributable to local auxin biosynthesis and polar transport, is largely responsible for lateral organ morphogenesis and the establishment of plant architecture. Here, we show that three Arabidopsis INDETERMINATE DOMAIN (IDD) transcription factors, IDD14, IDD15, and IDD16, cooperatively regulate auxin biosynthesis and transport and thus aerial organ morphogenesis and gravitropic responses. Gain-of-function of each IDD gene in Arabidopsis results in small and transversally down-curled leaves, whereas loss-of-function of these IDD genes causes pleiotropic phenotypes in aerial organs and defects in gravitropic responses, including altered leaf shape, flower development, fertility, and plant architecture. Further analyses indicate that these IDD genes regulate spatial auxin accumulation by directly targeting YUCCA5 (YUC5), TRYPTOPHAN AMINOTRANSFERASE of ARABIDOPSIS1 (TAA1), and PIN-FORMED1 (PIN1) to promote auxin biosynthesis and transport. Moreover, mutation or ectopic expression of YUC suppresses the organ morphogenic phenotype and partially restores the gravitropic responses in gain- or loss-of-function idd mutants, respectively. Taken together, our results reveal that a subfamily of IDD transcription factors plays a critical role in the regulation of spatial auxin accumulation, thereby controlling organ morphogenesis and gravitropic responses in plants.
An experiment was conducted to estimate by simple linear regression the levels of endogenous amino acids in digesta collected from the distal ileum in pigs. Six barrows, average initial BW 35 kg, were fitted with a simple T-cannula at the distal ileum and fed six diets according to a 6 x 6 Latin square design. Six cornstarch-based diets containing six levels of CP from soybean meal (4, 8, 12, 16, 20, and 24% CP, respectively) were formulated. Chromic oxide (.4%) was included as the digestibility marker. Each experimental period consisted of 8 d. Ileal digesta were collected, at 2-h intervals, for a total of 24 h during d 7 and 8. There were linear relationships (P < .001) between dietary contents of apparent ileal digestible and total amino acids, irrespective of the ranges in graded dietary levels of amino acids. Determined with the regression technique, the endogenous levels of the indispensable amino acids (grams/kilogram of DMI) were as follows: arginine, .64; histidine, .23; isoleucine, .46; leucine, .69; lysine, .47; methionine, .13; phenylalanine, .31; threonine, .69; and valine, .54. Differences in the ranges of graded dietary levels of amino acids resulted in large differences in the estimated amounts of endogenous amino acids in ileal digesta. Furthermore, it seems that the levels of endogenous amino acids, as grams/kilogram of DMI, were constant at different dietary levels of amino acids, whereas the contributions of endogenous amino acids, as percentages of their dietary contents, decreased curvilinearly with increasing dietary contents. Therefore, apparent ileal digestibilities of amino acids were quadratically related to their dietary contents until plateau digestibilities were reached, whereas the true ileal digestibilities of amino acids were independent of their respective dietary contents. Furthermore, true ileal amino acid digestibilities should be determined from their corresponding plateau apparent ileal digestibilities. In conclusion, the levels of endogenous amino acids in ileal digesta can be determined reliably from the linear relationships between dietary contents of apparent ileal digestible and total amino acids. An important methodological consideration in the determination of endogenous amino acids by regression analysis is to design an appropriate range of graded dietary levels of amino acids.
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